BRPI0721538A2 - Methods of treating pest infestation, killing invertebrate pest, inhibiting remodeling event and incubating eggs in non-ectoparasitic invertebrate in invertebrate population, controlling or killing invertebrate pest population, and selecting agent chelator as inhibitor candidate of invertebrate remodeling event from collection of metal chelating agents - Google Patents

Abstract

Methods of Treatment of Plague Infestation, Invertebrate Plague Extermination, Remodeling Event Inhibition and Egg Incubation in Non-Ectoparasitic Invertebrate Population, Control or Extermination of Invertebrate Plague Population and Agent Selection Chelating As Candidate Inhibitor of Invertebrate Remodeling Event from Metal Chelating Agents Collection. The present invention is directed to methods of treating plague infestation by inhibiting plague metabolic processes, such as, for example, processes involved in invertebrate remodeling.

Description

"Methods of Treatment of Pest Infestation, Invertebrate Plague Extermination, Remodeling Event Inhibition and Egg Incubation in Non-Ectoparasitic Invertebrate Population, Control or Extermination of Invertebrate Plague Population and Selection of Chelating Agent As Event Inhibitor Candidate

*

^ Invertebrate Remodeling from Collection

«T

of Metal Chelating Agents "

Descriptive Report

Field of the Invention

The present invention relates to compositions and methods for inhibiting the activity of an enzyme or enzymes that are directly or indirectly involved in invertebrate remodeling events. In particular, the invention relates to compositions and methods for controlling invertebrate multicellular organisms having cross-linked protein structures that include, but are not limited to, eggs, shells, carapaces, exoskeletons, cysts, cocoons or ootheca. . The invention also provides methods for inhibiting processes such as pololysis, ecdysis, egg hatching, encystation, shedding and metamorphosis. The invention also provides methods and compositions for preventing, treating or controlling infestations of an invertebrate pest undergoing remodeling events.

Background of the Invention

Pests that undergo remodeling events, such as egg hatching, moulting and / or adult pupal metamorphosis, cause significant problems in a wide variety of situations. For example, pests undergoing such remodeling events may externally infest humans or animals and irritate, injure and / or cause infestations, particularly in domesticated humans and animals. These pests can also internally infest humans and animals, causing infection, gastrointestinal problems, bloating, and / or lymphatic problems and blood loss. Pests undergoing remodeling events may also infest plants and their larvae or other life-cycle stages may eat leaves, flowers, roots and fruits causing significant damage to commercially important crops. Other pests that undergo these remodeling events infest the environment and cause human or animal disease or property damage. For example, termites cause significant property damage and the presence of dust or mites can cause asthma in humans.

A large number of pesticides are known to control or eliminate pests in plants, humans, animals and the environment. These pesticides may be used in the form of aerosols, space spray, liquids, soaps, shampoo, wettable talc, granules, bait, powders, tablets and the like.

Conventional pest control methods depend on the use of chemical pesticides such as chlorinated hydrocarbons (DDT, endosulfan, etc.), synthetic and natural pyrethrins (pyrethrin, permethrin, cypermethrin, deltamethrin), known insect growth regulators. because it interferes with the synthesis of chitin, insecticidal bacterial toxins (Bacillus thuringiensis (Bt) toxins) and nematicides, including both fumigants and non-fumigants (ie formulated granules or liquids). However, significant problems are associated with pesticide use, including target organ commonality and modes of action leading to the development of pest resistance, the need for increased pesticide use, the persistence of chemicals in the environment. and in plant and animal tissues, detrimental effects on the host and non-target organisms and lack of ovicidal activity.

The modern approach to pest species control depends on a combination of factors, including the use of appropriate management strategies that aim to minimize pesticide use but still allow for effective control. This change in control approach has been necessary due to overuse and reliance on chemicals leading to greater resistance problems to many commonly used chemicals. Moreover, due to their often quite specific modes of action, a number of chemicals used in the area often only aim at specific stages of the life cycle when the appropriate target is being expressed. For example, a pesticide can control pests by killing larvae only after they emerge from eggs, or kill the pest during their pupal or adult life stages. However, any eggs present in pesticide application are often unaffected and with maturity and hatching result in reinfestation of the plant, human, animal or environment. This results in repeated pesticide application or prolonged pesticide exposure being required for continued pest control. This is not only inconvenient and costly, but also increases the risks to the environment, plant, human or animal.

Accordingly, there remains a need to provide alternative methods and compositions that are effective in preventing or controlling remodeling events associated with pest development through all different stages of development to provide more efficient and effective control. Summary of the Invention

The present invention is directed to methods of treating pest infestation by inhibiting pest metabolic processes, such as, for example, processes involved in invertebrate remodeling. In specific embodiments, the methods of the invention comprise reducing shedding of an invertebrate external contact of a pest with a compound of formula (!):

wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (R 5) 2 -Z-C (R 5) 2-;

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, alkyl SO 3 C 1-6, NH 2, alkyl NHC 1 - e or N (C 1-6 alkyl) 2, or R 1 and R 19 taken together are -C (R 5) 2 -, -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol) halogen , CN, C (R 6) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) 6) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO 3 H, alkyl SO 3 Cl 1-6, NH 2, alkyl NHC 1-6 or N (C 1-6 alkyl) 2;

each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or decrease the shedding rate of said invertebrate.

Other embodiments of the invention comprise pest infestation treatment methods comprising decreasing an invertebrate encystation by external contact of a pest with a compound of formula:

wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (R 5) 2 -Z-C (R 5) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, COC 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NII 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, "10 CN, C (R 6) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, C03 -C 6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring;

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C 2 -C 6 alkyl -6, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2;

each R6 is independently selected from hydrogen and halogen.

nio; and

Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the encystation rate of said invertebrate.

The invention further contemplates methods of treating pest infestation comprising decreasing invertebrate apolysis by external contact of a pest with a compound of formula:

wherein X is selected from a covalent bond -C (R 5) 2-, -Z- or -C (RS) 2 -Z-C (RS) 2-;

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R5) 3, CO2H, C1 -C6 alkyl, SO3H, alkyl SO3 C1-6 alkyl, NH2, alkyl NHC1-6 or N (C1-6 alkyl) 2, or R1 and R19 taken together are -C (R5) 2- -C (R 5) 2 -C (R 5) 2-, -CR 5 -CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-5 alkyl) 2, - CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO 3 H, alkyl SO 3 Cl 1-6, NH 2, alkyl NHC 1-6 or N (C 1-6 alkyl)

each R6 is independently selected from hydrogen and halogen.

nio; and

Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) -

and -C (S) -;

a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the rate of said invertebrate polysis.

In yet another alternative embodiment, the methods of the invention

involve treating pest infestation comprising inhibiting metamorphosis of an invertebrate by external contact of said pest with a compound of formula:

where X is selected from a covalent bond -C (R5) 2-, -Z- or

-C (R5) 2 -Z-C (R5) 2-;

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 5) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, alkyl SO 3 C 1-6, NH 2, alkyl NHC 1 - e or N (C 1-6 alkyl) 2, or R 1 and R 19 taken together are -C (R 5) 2- -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, C 3 -C 6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO2H), NH (C1-6 alkylene) N (C1-0 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, C02C1 alkyl -6, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2;

each R6 is independently selected from hydrogen and halogen.

nio; and

Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the rate of metamorphosis of said invertebrate.

In the methods of the invention, It is preferred that the compound is a metal chelating agent, wherein the metal chelating agent has at least two polar atoms capable of coordinating simultaneously with a metal ion, has a clogP value of / 1 and <4. ; and / or a molar refractivity in the range of 40 to 90 cm 3 / mole. In specific embodiments, the metal chelating agent is not 1,10-phenanthroline. In other embodiments, the chelating agent is not a dipyridyl compound.

The methods of the invention contemplate the use of multiple pesticides and interventions to treat infestations. In specific embodiments, the methods further comprise contacting said pest with a second pesticide.

Other aspects of the invention include methods of treating pest infestation comprising reducing shedding of an invertebrate by external contact of a pest with a compound of formula (II):

(II)

10

where Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1 -C 6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen CN, C (R 2g) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NII 2, NHC 1 alkyl, or N (C 1-6 alkyl) 2; or

R 25 and R 26 together with the carbon atoms to which they are attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or decrease the shedding rate of said invertebrate.

Still another embodiment of the invention discloses a method of treating pest infestation comprising decreasing an invertebrate encystation by external contact of the pest with a compound of formula (II): wherein Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R 2 O) S, CO 2 H, C 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1 -C 6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen CN, C (R2g) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or

R 25 and R 26 together with the carbon atoms to which they are attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

20

R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the encystation rate of said invertebrate.

The invention also contemplates methods of treating pest infestation comprising decreasing invertebrate apolysis by external contact of a pest with a compound of formula (II):

15

(Π)

where Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R29) 3, CO2 H, C1 -C6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1- alkyl, N (C1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen CN, C (R 29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-5 alkyl) 2; or

R 25 and R 26 together with the carbon atoms to which they are attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, a C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or decrease the rate of said invertebrate polysis. Still another alternative method of treating pest infestation comprises inhibiting metamorphosis of an invertebrate by external contact of said pest with a compound of formula:

where Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6 halogen CN,

C (R 29) 3, CO 2 H, COC 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl, N (C 1-0 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R25 and R26 are independently selected from hydrogen,

C 1-6 kilogram, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 2g) 3, CO2H, C1 -C6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-10 alkyl) 2; or

R25 and R26 together with the carbon atoms to which they are

so attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, n (r30 ) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, a C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the rate of metamorphosis of said invertebrate.

Again, in each of the preceding methods, the compound is

preferably a metal chelating agent, wherein the metal chelating agent has at least two polar atoms capable of coordinating simultaneously with a metal ion, has a clogP value of / 1 and <4; and / or and a molar refractivity in the range of 40 to 90 cm 3 / mole. However, it is contemplated that the metal chelating agent is not 1,10-phenanthroline. The foregoing methods may further comprise contacting the pest with a second, third, fourth or more pesticides. In addition, the pest may be treated multiple times with the various pesticides described herein.

In preferred embodiments of the invention, the methods described

in this document produce a greater decrease in sheath shedding, encystation, apolysis, or metamorphosis than is observed with the administration of 1.10 phenanthroline. In particularly preferred embodiments, the methods are employed to kill an invertebrate pest, said method comprises external contact of said pest with a compound of formula (I):

wherein X is selected from a covalent bond, -C (R5) 2-, -Z- or

-C (RS) 2 -Z-C (RS) 2-;

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, alkyl SO 3 C 1-6, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, or R 1 and R 19 taken together are -C (R 5) 2- -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, CN, C (RS) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, C03 -C 6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring; each R5 is independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO 3 H, alkyl SO 3 Cl 1-6, NH 2, alkyl NHC 1-6 or N (C 1-6 alkyl) 2;

each R6 is independently selected from hydrogen and halogen.

nio; and

Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to kill said invertebrate.

Other preferred embodiments are directed to methods for killing an invertebrate pest, said method comprising external contact of said pest with a compound of formula (II):

15

where Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R29) s, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1 -C 6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R25 and R26 are independently selected from hydrogen,

C 1-6 kilogram, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, C 1 -C 6 alkyl, SO 3 H, C 1 -C 6 alkyl, NH 2, C 1 -C 6 alkyl or N (C 1 -C 6 alkyl) 2; or

R25 and R26 together with the carbon atoms to which they are

so attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R30 is independently selected from hydrogen, alkyl

C 1-6, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to kill said invertebrate.

In the killing methods of the invention, the invertebrate pest is selected from the group consisting of nematodes, trematodes, basketids, lice, fleas, mites and scabies, moths, beetles, caterpillars, butterflies, termites, arachnids, cockroaches, centipedes, fleas and mites.

It is preferable that the methods be such that they are used to kill at least some of the invertebrate pests that are infecting a host. In preferred embodiments, the methods yield results in which at least 25% of the pests in a given infestation are killed. In other modalities, at least 30% of the pest population is killed. In yet another alternative embodiment, at least 50% of the invertebrate population in a given infestation is killed. In yet another preferred embodiment, at least 75% of the invertebrate pest population in a given infestation is killed.

Also contemplated are methods of inhibiting a remodeling event in an invertebrate population comprising contacting said invertebrate population with a compound of formula (I):

wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (RS) 2 -Z-C (RS) 2-;

20

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, COC 1-6 alkyl, SO 3 H, alkyl SO 3 C 1-6, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, or R 1 and R 19 taken together are -C (R 5) 2- -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, C 3 -C 6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together

with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, C02C1 alkyl -6, SO3H, alkylSO3 Cl-6, NH2, alkyl NHCl-β or N (C1-6 alkyl) 2;

each R 6 is independently selected from hydrogen and halogen; and

Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said invertebrate remodeling event, wherein said invertebrate remodeling event is not egg hatching and said invertebrate is not an ectoparasite insect.

Another method of the invention is to inhibit a remodeling event in an invertebrate population comprising the internal contact of said invertebrate population with a compound of formula.

wherein Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R29) S, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1 -C 6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen CN, C (R 29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-0 alkyl) 2; or

(II):

(H)

R 25 and R 26 together with the carbon atoms to which they are attached form a 5- to 6-membered heterocyclic or carbocyclic ring; R27 is C1-6 alkyl, C1-6 branched chain alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) S, N (R30) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said invertebrate remodeling event, wherein said invertebrate remodeling event is not egg hatching and said invertebrate is not an ectoparasite insect.

The invention also provides methods of inhibiting egg hatching in a non-ectoparasite invertebrate an invertebrate population comprising contacting said invertebrate with a compound of formula (I): wherein X is selected from a covalent bond, -C (R 5 ) 2-, -Z- or -C (RS) 2-ZC (RS) 2-;

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NH 1 -C 5 alkyl or N (C 1-6 alkyl) 2, or R 1 and R 6 taken together are -C (R 5) 2 -, -C (RS) 2 -C (RS) 2-, -CRS = CRS-, C (O), C (S) or NH;

"R2, R2 ', R3, R3', R4 and R4 'are independently selected from

hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, halogen, CN, C (R6) 3, CO2H, CO2C1-6 alkyl, SO3H, SO3C1-6 alkyl, NII2, NHC1-6 alkyl or N (C1-6 alkyl) 2, -CH2CHNH (CO2H), NH (C1-6 alkylene) N (C1-6 alkyl) ) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together

with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, C1-6 branched chain alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO3H, alkyl SO3 -C6, NH2, alkyl NHC1- e or N (C1-5 alkyl) 2;

each R 6 is independently selected from hydrogen and halogen; and

Z is selected from a covalent bond, -NI-I-, -O-, -S-, -C (O) - and -C (S) -;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said egg hatching.

Also provided is a method of inhibiting egg hatching in a non-ectoparasite invertebrate in an invertebrate population comprising contacting said invertebrate with a compound of formula (II):

where Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, C 3 -C 6 alkyl NH 2, NHC 1 -C 6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or

R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen CN, C (R 29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, C 1 -C 6 alkyl, NH 2, N 1 -C 1 alkyl or N (C 1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 to 5 membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (RSO) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said egg hatching.

Preferably, the non-ectoparasite invertebrate is selected from the group consisting of nematodes, trematodes and basketids. In more preferred embodiments, the invertebrate is a nematode. Preferably, the nematode is inhibited at its larval stage.

The present invention provides additional methods for identifying and selecting a chelating agent as a candidate invertebrate remodeling event inhibitor from a collection of metal chelating agents;

said method comprising selecting a chelating agent of

metal having at least two polar atoms capable of simultaneously coordinating with a metal ion and

(i) a clogP value of / 1 and <4; and / or

ii) a molar refractivity in the range of 40 to 90 cm3 / mole.

Accordingly, the methods of the invention may be used to screen combinatorial libraries by rational drug model of agents that may be used as invertebrate remodeling inhibitors and / or as pesticides in general.

Still another aspect of the invention involves screening assays in which agents are identified and / or selected. Such methods involve identifying or selecting a chelating agent as a candidate inhibitor of invertebrate remodeling events from a collection of metal chelating agents of formula (II):

A: Yi Y2

where Y1 and Y2 are independently selected from O, NR28,

or S;

R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio and halogen, CN, C (R29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl, N (C1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen CN, C (R 29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1 alkyl, or N (C 1-0 alkyl) 2; or

R 25 and R 26 together with the carbon atoms to which they are attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (Rao) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof;

said method comprising selecting a metal chelating agent of formula (II) having at least two polar atoms capable of simultaneously coordinating with a metal ion and i) a clogP value of / 1 and <4; and / or

ii) a molar refractivity in the range of 40 to 90 cm3 / mole.

In yet another alternative embodiment, the methods of the invention involve screening methods for identifying or selecting a chelating agent as a candidate invertebrate remodeling event inhibitor from a collection of formula I metal chelating agents:

wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (R 5) 2 -Z-C (R 5) 2-;

R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, alkyl SO 3 C 1-6, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, or R 1 and R 19 taken together are -C (R 5) 2- -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, C03 -C 6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO3 H, alkyl SO3 Cl-6, NI-12, alkyl NHCl-e or N (C1-6 alkyl) 2;

each R6 is independently selected from hydrogen and halogen.

nio; and

Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

said method comprising selecting a metal chelating agent of formula (I) having at least two polar atoms capable of simultaneously coordinating with a metal ion and

(i) a clogP value of / 1 and <4; and / or ii) a molar refractivity in the range of 40 to 90

cm 3 / mole.

The screening assays of the invention may be combined with conventional biological assays employed to determine the effectiveness of a given agent as a pesticide. As such, the above screening assays may be combined with assays designed to determine the effect on egg hatching, molting, metamorphosis and the like, as well as in vitro enzyme assays that determine the activity of one or more of the enzymes involved in a or more of the remodeling events. In specific embodiments, screening assays may be combined with assays that determine the efficacy of compounds as protease inhibitors and the like.

Brief Description of the Figures Figure 1: presents a SDS-PAGE analysis with wash substrate of protease activity obtained from various hair and lice egg samples (ESW eggshell washes) following gel staining as Coomassie and discoloration. Column 1 shows the protease activity detected in washes obtained from un hatched lice eggs within 12 hours of hatching (sample 1) in the highest molecular weight region of the SDS gel above 50kDa (Figure IA, column 1). A similar pattern of protease activity was detected in washes taken from human hair samples followed by removal of louse eggs (sample 2) (Figure IA, column 2). However, treating the hair with 1% sodium hypochlorite before collecting the washes (sample 3) completely removed protease activity (Figure IA, column 3). Hypochlorite treatment was also able to remove foreign proteases from un hatched louse eggs (sample 4) (Figure IA, column 4). Hypochlorite was used to treat un hatched eggs prior to ESW collection for all subsequent protease analyzes.

Several distinct proteases were observed in the ESWs of hypochlorite-treated eggs collected within 2 hours of hatching (sample 5) (Figure 1B). Protease activity ranges were detected around 25-30 kDa, 50 kDa and there were a number of lighter ranges detected above 50 kDa. These proteases were specifically associated with lice eggs at the time of hatching and were called eggshell washes (ESW).

Figure 2: Proteases present in louse ESWs have been further

characterized by their mechanistic class. Incubation with the metal chelating agents EDTA and 1,10-phenanthroline to inhibit metalloproteins resulted in a reduction in protease activity compared to untreated controls (Figure 2A and 2B, respectively). In contrast, there was no apparent reduction in protease activity when ESW was incubated with the serine / cysteine protease inhibitor PMSF (Figure 2B), cysteine protease inhibitor E-64 (Figure 2B) or pepstatin aspartic protease (data not shown).

Figure 3 shows a two-dimensional gelatin SDS-PAGE that was used to evaluate the number of protease species present in louse ESWs. Each of the three major regions of protease activity in the one-dimensional gelatin SDS-PAGE (Figure 1B) determined for a number of distinct proteases present in louse ESWs with activity in the 25-30 kDa molecular weight range determined for at least seven distinct proteases with isoelectric points in the neutral to alkaline pH range, while the protease activity range of about 50 kDa is determined for at least eleven distinct protease regions with isoelectric points in the acid to neutral pH region. At least five proteases with molecular weights above 75 kDa were also observed.

In order to further investigate the effect of 1,10-phenanthroline on ESWs protease activity, the proteases were separated by two-dimensional gel electrophoresis and the gel incubated in the presence of 10 mM 1,10-phenanthroline. The results of these studies confirmed the inhibitory effect of this metalloprotease inhibitor on louse egg protease activity. There was a general reduction in protease activity in the 25-30 kDa region and a clear reduction in proteases present around the 50kDa region and above 75 kDa (Figure 3B).

Figure 4: shows the effect of 1,10-phenanthroline on hatching of

egg in lice. Eggs were treated 5 days after laying and then hatching was observed over time.

Figure 5: shows the effect of Lannate® containing metomil on egg hatching in Helicoverpa eggs. The efficacy of ovicide was evaluated at 5 mM, 2.5 mM, 2.25 mM and 0.125 mM metomil.

Figure 6: shows the effect of 2-acetyl-1-tetralone on egg hatching in Helicoverpa armigera eggs. Ovicidal efficacy was evaluated at 2mM and ImM of 2-acetyl-1-tetralone.

Figure 7: shows the effect of 2-acetyl-1-tetralone on the outbreak of

eggs in Plutella eggs. Ovicidal activity was evaluated at 2mM, ImM, 0.5mM and 0.1mM.

Figure 8: shows the effect of 5,5'-dimethyl-2,2'-dipyridyl on egg hatching in H. contortus eggs. Ovicidal efficacy was evaluated - 10 at 180 µg / mL, 18 µg / mL, 1.8 µg / mL and 0.18 µg / mL.

Figure 9: presents the effect of ivermectin on egg hatching in II eggs. contortus The effectiveness of ovicide was evaluated at 200 μg / mL, 100 μg / mL, 50 μg / mL, 25 μg / mL and 12.5 μ§ / ηι] ϋ.

Figure 10: shows the effect of 2-acetyl-1-tetralone on egg hatching in H. contortus eggs. The effectiveness of ovicide was evaluated at 110 μg / mL and 22 μg / mL.

Description of the Invention

In one aspect of the invention there is provided a method of inhibiting a remodeling process in an invertebrate by externally applying a pesticidal composition substantially as described hereinbelow. The remodeling process to be inhibited can be any process that is involved in the life cycle of an invertebrate pest. As such, the invention contemplates inhibiting processes such as egg hatching, encysting, shedding, pololysis, ecdysis or metamorphosis. Without being limited to a given theory or mechanism of action, the invention may, but need not necessarily, involve the inhibition of a protease enzyme involved in this remodeling process. Protease enzymes are known, for example, to be involved in protein hydrolysis in eggs, sheaths, carapaces, exoskeletons, cysts, cocoons or ootheca, weakening the structure and at least partially allowing the invertebrates to break free of the structure. In some alternative methods of the invention, it is contemplated that the remodeling process indirectly involves a protease enzyme, for example, a given protein or peptide may be required for the remodeling process, such as a hormone that signals that The remodeling process must take place and the compositions of the invention are capable of inhibiting the production of this hormone. Preferably, the protease enzyme is a metalloprotease enzyme.

The term "remodeling event" refers to an event in the life cycle of an invertebrate that alters the immediate environment of invertebrates or alters the physical shape of invertebrates and facilitates the organism's progression from one life stage to the next. life to the next stage of life. Examples of remodeling events include egg hatching, encystment, cuticle or exoskeleton apolysis, cuticle or exoskeleton ecdysis, and metamorphosis.

As used herein, "egg hatching" refers to hatching of a thin-membrane egg invertebrate where hatching is aided by protease enzymes. Thin-membrane eggs include those eggs which have shells or cuticles predominantly comprising a protein matrix, with or without tanned protein, are gas permeable, but essentially water impermeable, are generally non-mineralized, are less than 20 mm in length and cannot be hatched solely by mechanical means, for example by unassisted chewing or rupture.

As used herein, "encystment" refers to the emergence of an embryonic larval or quiescent protozoa, taeniid, trematode or other species from a closed membranous sac or tissue cavity at some stage in its life cycle. These bags may be formed in part or entirely from proteinaceous material that must be reshaped in order for the invertebrate to emerge. Cysts may be formed within the body of a host, such as a human or a snail, or directly deposited in the environment. Inquisition must occur at the right time and in the right way in order for the invertebrate to continue with subsequent stages of its life cycle.

As used herein, "sheath shedding" refers to a process of moulting on worms such as nematodes. After hatching, a nematode goes through four larval stages before emerging as an immature adult. Larval stages are coated in a cuticle or sheath that has a protective role. The process involves two steps, the synthesis of a new cuticle and shedding or peeling of the old cuticle. Sheath shedding may also be essential to allow a new host to become infected. For example, shedding of H. contortus third larval stage sheath into the rumen of a host results in host infection. Environmental conditions in the rumen activate secretory cells in the nematode to release hormones. Hormones act on excretory cells and stimulate water uptake, which in turn activates enzymes that are released into the space between the new and old cuticles. Enzymes weaken the old cuticle, which then breaks down, allowing the worm to break free of the old cuticle.

As used herein, "apolysis" refers to the separation of the epidermis cuticle from an invertebrate. This separation allows the formation of a new cuticle without exposure to the environment. During this process, enzymes are secreted from the invertebrate, which digest the inner layers of the cuticle.

As used herein, the term "ecdise" refers to the dislocation of an old cuticle. Ecdysis occurs after the apolysis. After apolysis, mute fluid containing inactive enzymes is secreted into the space between the epidermis and the old cuticle. The new cuticle is then formed. Enzymes in the molting fluid are then activated and the lower regions of the old cuticle, the endocuticle and the mesocuticle, are digested. The exocuticle and epicuticle of the old cuticle, which are not digested, are then stripped.

As used herein, "metamorphosis" refers to the biological process in which some invertebrates, after hatching, undergo a conspicuous change in shape or structure through cell growth and differentiation, which is often accompanied by a change in habitat. and / or behavior. Metamorphosis usually proceeds in distinct stages, usually starting with a larva or pupa, optionally passing through a pupa, and results in an adult. Metamorphosis of a chrysalis, usually shaped like an adult, can be marked by the development of wings. In contrast, other invertebrates may have larvae that differ substantially from the adult and go through an inactive stage called the pupa, from which an adult emerges. Growth and metamorphosis are controlled by hormones produced by the invertebrate. A combination of hormones can be used, for example, to secrete ecdisone (a steroid) and the juvenile hormone allows for the moulting or growing of a chrysalis or larva without metamorphosis maturation for an adult. When the juvenile hormone is no longer produced, the metamorphosis proceeds.

The term "exposing an invertebrate" as used herein refers to the exposure of invertebrates to any part of their life cycle, including, but not limited to, an invertebrate egg, ootheca, cyst, chrysalis. invertebrate, an invertebrate larva, an invertebrate instar, an invertebrate pupa, and any juvenile or adult stage of an invertebrate. The term "in contact" as used herein may refer to an external contact of the pest with the composition of the invention. Alternatively, the pest may contact the composition of the invention because the pest has ingested the composition. In yet another alternative, the pest is in contact with the invention because the pest host has ingested or been in contact with the composition and by being in physical contact with the host, the pest or ingesting or coming in external contact with the compounds. of the invention. It is noted that the compositions and methods of the invention are employed to kill, inhibit or otherwise disrupt the life cycle stage that is exposed to the compositions of the invention. For example, where the composition is used to inhibit egg hatching, the composition is exposed directly to the invertebrate egg rather than to a different stage in the invertebrate life cycle. As such, in the methods of the invention, it is not necessary for the inhibitory compositions to be ingested by the host or even the pest in order to have their inhibitory effects as required, for example, in the methods described in US Patent 5,766,609 and 6,150,125. . Preferably, the compositions and methods of the present invention are such that the compositions are simply in contact with the outer surface or environment of the invertebrate and act either to kill the invertebrate directly, or to retard, inhibit or otherwise prevent the invertebrate. invertebrate progresses to the next stage of its life cycle. Accordingly, in certain embodiments, it is contemplated that the methods of the invention are used to treat a pest infestation by killing, inhibiting or otherwise undoing such an infestation by preventing and removing the infestation at the invertebrate life-cycle stage in which the disease occurs. composition is applied. Therefore, in the present invention, a method of treating flea infestation is provided in such a way that the flea does not necessarily ingest the compositions of the invention, whereas for nematodes and caterpillars the composition may be ingested. US 5,766,609, on the other hand, requires that a flea must ingest certain compounds in order to inhibit proteases that form significant components of the midgut of the flea and thus reduce flea fecundity by such ingestion of protease inhibitors. . Thus, a key difference between the use of the compositions of the present invention and the methods taught by US Patent 5,766,609 is that the compositions of the present invention act directly on the animal in contact with / fed with / otherwise exposed to the compositions, in instead of the compositions having an effect on subsequent generations (eg where flea eggs are exposed, there is a decrease in egg hatching while fleas may well remain unaffected).

The term "metal chelating agent" as used herein refers to a molecule having at least two polar atoms, such as nitrogen, oxygen, sulfur and phosphorus, and are situated in the molecule such that they are. able to simultaneously coordinate on a metal ion. The metal chelating agent also has a clogP value of / 1 and <4 and / or a molar refractivity in the range of 40 to 90 cm 3 / mole. In some embodiments, the chelating agent has a clogP value of / 1 and <3 and a molar refractivity in the range of 40 to 70 cm 3 / mole. In some embodiments, the chelating agent pLDso is / 2, preferably / 3, more preferably / 4. In some embodiments, the constant or LogKb association of zinc metal chelating agents is> 5.00.

Metal ions that are capable of being coordinated by the metal chelating agent are any metal ions that occur in metalloproteases, particularly metalloproteases that are involved in breaking down structures containing cross-linked proteins associated with eggs, sheaths, carapaces, cuticles. , exoskeletons, cysts or ootheca and / or that facilitate the progression of the organism from one life stage to the next life stage. Such metal ions include divalent and trivalent metal ions, particularly divalent alkaline earth metal ions and divalent or trivalent transition metal ions. In some embodiments, the coordinate metal ions are selected from Ca ++, Mg ++, Cu ++, Fe ++, Zn ++ and Fe +++, especially Cu ++, Fe ++ and Zn ++, more especially Zn ++.

ClogP is a calculated prediction of the IogP value of a compound. The IogP value of a compound, which is the logarithm of its partition coefficient between n-octanol and water [(log (C0ctanoi / Water)]), is a well-established measure of the hydrophilicity of the compound. in the atom type and includes information regarding various atomic properties such as atomic number, ring members, immediate neighbor bond types and aromatic state.ClogP can be calculated using a clogP program (Biobyte).

Molar refractivity is a measure of the volume occupied by an atom or group and depends on temperature, refractive index, pressure. Molar refractivity provides an indication of the size of the molecule and the polarizability of the molecule. Molar refractivity was calculated using the CMR (Calculated Molar Refractivity) module of the ClogP (Biobyte) computer program.

Log LD50 is obtained from the observed percentage of ovicidal activity by conversion using a modified logite transformation. The observed percentage values of ovicidal activity were transformed using the equation:

BA = Yog ((5 +%) / (105 -%))

The additional 5% was used to allow a number to be calculated when 0% and 100% activity were observed. BA is a crude correction for recording the concentration at which the compounds were tested. Correlation between BA and LD50 values allows the calculation of Log LD50. A pLDso value less than 2 is considered inactive (Class 0), a pLDso value between 2 and 3 is considered weakly active (Class 1), a pLDso value between 3 and 4 is considered moderately active (Class 2) and a pLDso greater than 4 is considered strongly active (Class 3). In preferred embodiments, the pLDso value is greater than 2, especially greater than 3, and more especially greater than 4.

Without wishing to be bound by theory, it is believed that

Metal chelators bind and remove the metal ions required for metalloprotease activity making the protease inactive. This theory is supported by the addition of metal ions reversing the inhibitory effect of the metal chelating agent.

»

In some embodiments, the metal chelating agent is a

post of formula (I):

wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (R 5) 2 -Z-C (R 5) 2-;

R1 and R1 'are independently selected from hydrogen, alkyl

C 1-6, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, C 2 -C 6 alkyl 6, SO 3 H, alkyl SO 3 Cl 1-6, NH 2, alkyl NHC 1-6 or N (C 1-8 alkyl) or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C (R 5) 2-, -CR5-CR5-, C (O), C (S) or ΝΗ;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, C03 -C 6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO2H), NII (C1-6 alkylene) N (C1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together

with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO3II, alkyl SO3 C1-6, NH2, alkyl NHCl1-6 or N (C1-6 alkyl) 2;

each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

or a pharmaceutically, veterinarily or agriculturally salt

acceptable to them.

Preferred compounds of formula (I) have at least one of the following characteristics:

R 1 and R 1 'are independently selected from C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, CO 2 H, C 2 -C 6 alkyl -6, SO3H, alkyl SO3 C1-6, NH2, alkyl NHC1- e or N (C1-6 alkyl) 2, more preferably hydrogen or C1-3 alkyl, most preferably hydrogen or methyl;

R2 and R2 'are independently hydrogen or C1-3 alkyl, more preferably hydrogen;

R3, R3 ', R4 and R4' are independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, C1-6 alkylthiol or C1-6 alkyl, preferably hydrogen or C1-3 alkyl, more preferably hydrogen or methyl; or R3 and R4 and / or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- to 6-membered carbocyclic or heterocyclic ring, preferably an aromatic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, C1-6 alkylthiol or CO2C1-6 alkyl, preferably hydrogen. nio or C 1-3 alkyl, more preferably hydrogen or methyl;

each R5 is independently hydrogen or fluorine, especially where each R6 is fluorine;

X is a covalent bond, -CH 2 -Z-CH 2 - or Z, preferably a covalent bond; and

Z is -NH-, -O- or -S-, preferably -NH-.

In some embodiments, the substituents R1, R2, R3, R4, R1 ', R2', R3 'and R4' are electron donors, or do not affect the electron density of the pyridyl ring.

Preferred compounds are biaryl compounds of formula (I):

20

wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (RS) 2 -Z-C (RS) 2-;

R 1 and R 19 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen C (R 6) 3, CO 2 H, C 2 -C 6 alkyl, SO 3 H, alkyl SO 3 C 1-6, NH 2, alkyl NHC 1 - e or N (C 0 -C 6 alkyl) 2;

R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy , thiol, C1-6 alkylthiol, halogen, CN, C (R6) 3, CO2H, CO2C1-6 alkyl, SO3H, SO3C1-6 alkyl, NII2, NIIC1-6 alkyl or N (C1-6 alkyl) 2, -CH2CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered heterocyclic or carbocyclic ring; or

R2 and R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered heterocyclic or carbocyclic ring;

each R5 is independently selected from hydrogen, C1-6 alkyl, C1-6 branched chain alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2C1 alkyl -6, SO3H, alkyl SO3 -C6, NH2, alkyl NHC1- e or N (C1-6 alkyl) 2;

each R5 is independently selected from hydrogen and halogen.

nio; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof.

Preferred compounds of formula (!) Include

2,2'-dipyridyl, 6,6'-dimethyl-2,2'-dipyridyl, 5,5'-dimethyl-2,2'-dipyridyl, 5,5'-diethyl-2,2'-dipyridyl, 4 , 4'-Dimethyl-2,2'-dipyridyl, 2- (2-pyridinyl) quinoline, 2,2-dipyridylamine, 2,2 ', 6,2' -pyridine

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof.

In some embodiments, the preferred metal chelating agent is a compound of formula II:

where Y1 and Y2 are independently selected from O, NR28,

or S;

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-alkylthio -6, halogen, CN, C (R29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NII 2, NHC 1 -C 6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring;

R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen , CN, C (R 29) 3, CO 2 H, C 1 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-0 alkyl) 2; or

R 25 and R 26 together with the carbon atoms to which they are attached form a 5- to 6-membered heterocyclic or carbocyclic ring;

R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2, or a 5- or 6-membered carbocyclic ring or heterocyclic ring;

1

R28 is hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl;

R29 is hydrogen or halogen; and

each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5-6 membered carbocyclic ring or heterocyclic ring;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof.

Preferred compounds of formula (II) have at least one of the following characteristics:

R21, R22, R23 and R24 are independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, halogen, NH2, NHC1-6 alkyl, N ( C1-6 alkyl) or CN, preferably hydrogen or C1-6 alkyl, a branched chain C1-6 alkyl, more preferably hydrogen or C1-3 alkyl, especially hydrogen or methyl;

R25 and R26 are independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, halogen, NH2, NH (C1-6 alkyl), C1-6 alkylphenyl. (6) 2 or CN, preferably hydrogen or C1-6 alkyl, a branched chain C1-6 alkyl, more preferably hydrogen or C1-3 alkyl, especially hydrogen or methyl; or

R 25 and R 26 taken together with the carbon atoms to which they are attached form a 5 to 6 membered heterocyclic or carbocyclic ring, especially a carbocyclic ring, more especially a 6 membered unsaturated carbocyclic ring;

R27 is C1-6 alkyl, a branched chain C1-6 alkyl, C1-6 alkenyl, C1-6 alkynyl, C (R29) 3, or a 6-membered carbocyclic or heterocyclic ring; especially C1-6 alkyl, a branched chain C1-6 alkyl, or a 5- to 6-membered carbocyclic ring, preferably a C1-3 alkyl or a 6-membered carbocyclic ring; especially methyl or phenyl;

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof.

Preferred compounds of formula (II) include: dibenzoylmethane,

benzoylacetone, and 2-acetyl-1-tetralone,

or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof.

As used herein, the term "alkyl" refers to a straight or branched chain saturated hydrocarbon group and may have a specified number of carbon atoms. For example, C1 -C6 as in "C1 -C6 alkyl" includes groups having 1, 2, 3, 4, 5 or 6 carbons in a straight or branched arrangement. Examples of suitable alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 4 -methylbutyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 5-methylpentyl, 2-ethylbutyl and 3-ethylbutyl. C 1-6 alkyl as used herein also includes branched chain C 1-6 alkyl.

As used herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon group having one or more double bonds between carbon atoms and may have a specified number of carbon atoms. For example, C 2 -C 6, as in "C 2 -C 6 alkenyl", includes groups having 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. Examples of suitable alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, pentenyl and hexenyl.

As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon group having one or more triple bonds between carbon atoms, and may have a specified number of carbon atoms. For example, C 2 -C 6, as in "C 2 -C 6 alkynyl", includes groups having 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. Examples of suitable alkynyl groups include, but are not limited to, ethinyl, propynyl, butinyl, pentinyl and hexinyl. As used herein, the term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

The terms "alkyloxy" or "alkoxy" as used herein represent an alkyl group, as defined above, attached by an oxygen bridge. Examples of suitable alkyloxy groups include, but are not limited to, methoxy, ethoxy, n-propyloxy, t-propyloxy, n-butyloxy, i-butyloxy, t-butyloxy, n-pentyloxy and n-hexyloxy.

The term "alkylthio" as used herein represents an alkyl group, as defined above, linked by a sulfur bridge, examples of suitable alkylthio groups include, but are not limited to, methylthio, ethylthio, propylthio, β-propylthio, butylthio t-butylthio, t-butylthio, pentylthio, hexylthio.

The term "carbocyclic ring" as used herein refers to a 3 to 10 membered ring or fused ring system in which all ring-forming atoms are carbon atoms. The C 3-10 carbocyclic ring may be saturated, unsaturated or aromatic. Examples of suitable carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, phenyl, naphthyl and tetrahydronaphthyl.

The term "heterocyclic ring" as used herein refers to

refers to a 3- to 10-membered ring or a fused ring system in which at least one of the ring-forming atoms is a heteroatom. Preferably, the heteroatom is selected from nitrogen, oxygen, sulfur and phosphorus. The C 3-10 heterocyclic ring may be saturated, unsaturated or aromatic. Examples of suitable heterocyclic rings include, but are not limited to, benzoimidazolyl, benzofuranyl, benzofuranyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinolinyl, furanyl, imidazoyl, indolyl, indolazinyl, indolazinyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrido-pyridinyl, pyridazinyl, pyridyl, pyrimidylazoline, pyrimidylazoline tetrazolyl, tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, azetidinyl, aziridinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinhydridihydrobihydrobihydrobihydride imidazolyl, dihydroindolyl, dihid roisooxazolila, dihidroisotiazolila, dihidrooxadia-zolila, dihidrooxazolila, dihidropirazinila, it dihidropirazoli-, dihidropiridi-nila, dihidropirimidinila, dihidropirrolila, dihidroquino- linila, dihydro-tetrazolila, dihidrotiadiazolila, dihidrotiazolila, dihidrotie- nila, dihydro-triazolila, dihidroazetidinila, metilenodioxibenzoíla, tetra- hydrofuranyl, and tetrahydrothienyl, and N-oxides thereof. Bonding of a heterocyclyl substituent may occur via a carbon atom or by a heteroatom.

As used herein, the term "aryl" is intended to mean any stable monocyclic or bicyclic carbon ring of up to 6 atoms in each ring, wherein at least one ring is aromatic. Examples of such aryl groups include, but are not limited to phenyl, naphthyl and tetrahydronaphthyl.

The term "heteroaryl" as used herein represents a stable bicyclic or monocyclic ring of up to 6 atoms in each ring, wherein at least one ring is aromatic and at least one ring contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S. Heteroaryl groups within the scope of this invention include, but are not limited to, acridinyl, carbazolyl, cinolinyl, quinoxalinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline.

The compounds of the invention may be in the form of pharmaceutically, veterinarily or agriculturally acceptable salts. Suitable pharmaceutically acceptable salts include, but are not limited to, pharmaceutically acceptable inorganic acid salts, such as hydrochloric, sulfuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids, or pharmaceutically acceptable organic acid salts. such as acetic, propionic, butyric, tartaric, maleic, hydroxalic, fumaric, maleic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulfonic, toluenesulfonic, benzenesulfonic acid, salicylic, , glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric.

Basic salts include, but are not limited to those formed with pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonia and alkylammonia.

Basic nitrogen-containing groups can be quartenized

with such agents as lower alkyl halide such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates, such as dimethyl and diethyl sulfate; and others.

It will also be recognized that many compounds of the invention have asymmetric centers and are thus capable of existing in more than one stereoisomeric form. The invention thus also relates to compounds in substantially pure isomeric form at one or more asymmetric centers, for example greater than about 90% ee, such as about 95% or 97% ee or greater than 99%. % ee, as well as mixtures, including racemic mixtures, thereof. Such isomers may be prepared by asymmetric synthesis, for example using chiral intermediates or by chiral resolution.

A number of metal chelating agents and metalloprotease inhibitors useful in the present invention may be obtained commercially from chemical companies. Those not commercially available may be synthesized from commercially available raw materials using reactions known to those skilled in the art.

For example, substituted 2,2'-bipyridyls and 1,10-phenanthrolines may be obtained from halogenated 2,2'-bipyridyls or 1,10-phenanthrolines. For example, 2,2'-bipyridine-6,6'-dicarboxylic acid can be obtained from 6,6'-dibromo-2,2'-dipyridyl by exchanging metal halogen for butyl lithium by treatment with dry ice. and acidification [Buhleier et. al., Chem. Ber., 1978, 111: 200-204], Monosubstitution of a bipyridyl, for example, with CH 2 CHNI-12 (CO 2 H) at position 6, can be obtained by treating 6-methyl-2,2'- bipyridyl with N-bromosuccinimide, followed by alkylation with N-protected glycine ester. The protecting groups may then be removed by acid hydrolysis (Imperiali B. and Fisher S.L., J. Org. Chem., 1992, 57: 757-759).

2,2'-Dipyridyls may undergo nucleophilic substitution at positions C6 and C4 to introduce substituents. This reaction is most favorable when a halogenated dipyridyl is used as the raw material. For example, an amine may be introduced at C 6 and / or C 6 'using 6-mono or dihalogenated 2,2'-dipyridyl and reacting this feedstock with ammonia.

Bipyridyl sulfonic acids can be prepared from 2,2'-bipyridyl by heating with either oleum (a solution of sulfur trioxide in concentrated sulfuric acid) or mercury (I-I) sulfate / concentrated sulfuric acid at 300 ° C.

Asymmetrically substituted bipyridyls may be obtained from symmetrical bipyridyls, for example 6'-methyl-2,2'-bipyridyl-6-carboxylic acid may be prepared from oxidation by 6,6'-dimethyl-2,2'-bipyridyl selenium dioxide, followed by silver nitrate treatment (Al-Saya et al., European J. Org. Chem., 2004, 173- 182).

Compounds of formula (II) may be prepared by reacting an appropriately substituted benzaldehyde and a ketone such as acetophenone or acetone using an aldol reaction then converting the resulting hydroxy ketone to a 1,3-diketone as shown in Scheme 1:

1. Cli

2. catalysod hydrdysis base

Scheme 1

10

Compounds of formula (II) in the form of tetralones may be prepared using the Haworth reaction followed by substitution of a, as shown in Scheme 2: R-:

THE

u

η

π

1. AlCIj

2. ZtVHC

ο

3. H2SOi Uv

K--

THE

base

THE

H'

, Jt ν

Invertebrates that are inhibited from remodeling events in the present invention are pests that internally and externally infest humans or animals, infest plants or infest property or a particular environment. For example, pests that internally infest humans or animals include, but are not limited to, nematodes, trematodes and basketids; pests that infest humans or animals include, but are not limited to, lice, fleas, mites and scabies, pests plant-infesting include, but are not limited to, moths, beetles, butterfly caterpillars and nematodes, property-damaging pests include, but are not limited to, termites and pests infesting an environment include, but are not limited to, arachnids, cockroaches, centipedes, fleas and lice.

In another embodiment, a method of treating or preventing a pest infestation of a host or environment is provided comprising applying or administering to the host or environment an effective amount of at least one metal chelating agent, wherein the metal chelating agent has at least two polar atoms capable of simultaneously coordinating with a metal ion and

(i) a clogP value of / 1 and <4; and / or

ii) a molar refractivity in the range of 40 to 90 cm3 / mole;

or a pharmaceutically, veterinarily or agriculturally salt

acceptable to them.

The host treated by the methods of the invention may be selected from, but not limited to, the group consisting of humans, sheep, cattle, horses, pigs, poultry, dogs and cats. The treatment or prevention methods of the present invention may be applicable to plants and or other breeding, feeding or

«

housing of plagas. Plants treated by the methods of the invention are preferably selected from, but are not limited to, the group consisting of cotton, oilseeds or grain crops such as canola, forestry, such as trees, plant specimens such as trees, plants ornamentals such as shrubs, flowers such as chrysanthemum, michaelmas daisy, geraniums and roses, fruit trees such as apple, pear, plum, kiwi trees and citrus varieties, for example lemons, oranges, limes and grapefruits, cereals such as maize and sweet corn, vine crops such as grapes, tubers, grazing plants such as meadows and white clover, lucerne and lupins, and vegetables such as cabbage cultivation, for example , broccoli and cauliflower, cabbage, tomatoes, zucchini, leeks, lettuce and beans, as well as legumes such as beans, soybeans, guandu and chickpeas, and vine crops such as grape.

The environment to be treated by the methods of the present invention includes the environments of an animal, human or plant that is or may become pest-infested and includes, but is not limited to, surrounding plant, garden, grass, kennel soils. , barns or enclosure of animals, carpets, clothing, bedding and pest breeding sites. The environment also includes property that may be damaged by a pest, for example buildings, furniture and wood products, which may be damaged or destroyed by termites.

Preferred pests that undergo remodeling events and can be controlled by the methods described include, but are not limited to, species of a class, subclass or selected order of the phylum Platelminths, such as the Cestoda and Trematoda classes of the phylum Nemato, such as the Adenophoria or Secernentea classes of the phylum Arthropoda, as well as the classes Crustacea1 Arachnida, Insecta and Aearina.

There are two orders of the class and the other, namely, Cestodana and Euestodia, of which cyclophillideans are of utmost importance to humans because they infect people and livestock. Two important solitaires are the pig solitaire, Taenia soliurn, and the ox solitaire, T. saginata.

Of the trematode class, the Digenea subclass, which includes the trematodes. Trematodes can be classified into two groups, at the base of the system they infect. Tissue trematodes are species that infect the bile ducts, lungs, or other biological tissues, including pulmonary trematodes, Paragonimus westermani, and hepatic trematodes, Clonorchis sinensis, Fasciola hepatiea and Faseiola gigantiea. The other group is known as blood trematodes, and inhabit the blood at some stages of its life cycle. Blood trematodes include several species of the genus Schistosoma.

Nematodes, usually parasitic in humans, include tricabris, hookworms, oxyurids, mites, and branchias. Within the nematode phylum is the Adenophoria class and the Enoplia subclass, which includes the lower back. Most nematodes in this subclass are free-living, but the group also includes the order Trichiurida, which includes parasitic trichuris and triehina worms.

Also within the nematodes is the Secementea, subclass Rhabditia, which is mostly composed of parasitic nematodes, although there are some free-living species as well. An important order is Ascaridide, which includes worms that infect many terrestrial and marine mammals. Important families within this order include Ascarididae, which includes the giant intestinal roundworm and related species, and Toxocaridae, which includes canine, feline, and raccoon parasites, but which can be wildly parasitic in humans and cause visceral larva migrans. Another important order is Strongylida, which includes the genus Metastrongylus, a nematode of the Metastrongylidae family, commonly found as lungworms in pigs and sometimes causing verminous bronchitis. The subfamily Strongylinae (large strongyles) and Cyathostominae, (small strongyles) are important horse nematodes, while the Triehostrongylid family contains a number of economically important intestinal parasites of sheep and cattle, including Haernonchus contortus, H. placei. , Teladorsagia circumcincta, Trichostrongylus colubriformis and Cooperia spp.

In addition to mammalian host nematode parasites, there are several groups of plant parasitic nematodes that can cause various crop losses. The most common genera are Apheplochoid (leaf nematodes), Meloidogyne (gall nematodes), Heterodera, Globodera (cyst nematodes), such as potato nematodes, Nacobhus, Pratylenehus (lesion nematodes), Dityleneor Longusphus, Xiphinemaus, . Several species of plant parasitic nematodes cause histological damage to roots, including the formation of visible galls (Meloidgyne), which are useful signals for their field diagnosis. Some nematode species transmit plant viruses by their feeding activity in the roots. One such nematode is Xiphinema index, a vector of the GFLV (Grapevine Fanleaf Virus), an important grape disease. Other nematodes bind tree bark and forest trees. The most important representative of this group is Bursaphelenchus xylophilus, the pine nematode, present in Asia and America and recently discovered in Europe.

Of the crustacea, class Maxillopoda, subclass Maxillopoda, which includes fish louse.

From the Insecta, the orders include: Lepidoptera, Iememtera, Orthoptera, Psocoptera, Isoptera, Coleoptera, Dictyoptera, Thysanoptera, Homopolis, Diptera, Siphonaptera and Phthiraptera, which comprises Anoplura and Mallophaga.

Suitable pests that can be controlled using the methods of the present invention include:

(a) of the order Lepidoptera (Lepidoptera), for example, Adoxophysorana, Agrotis ypsilon, Agrotis segetum, Alabama argillacea, Antichesia gemmatalis, Argyresthia conjugella, Autographa gamma, Cacoecia murinana, Capua reticulana, Chonstoneura fumiferana , Choristoneura oceidentalis, Chrysodexis spp., Cirphis unipuneta, Cnaphalocrocis medinalis, Crocidolomia hinotalis, Crocidolomia pavonana, Cydia pomonella, Dendrolimus pini, Diaphania nitidalis, Diatraea granidiosella, Eariaspulanus, Elasopanus epiphytes Eupoecilia ambiguella, Feltia subterranean, Grapholitha funebrana, Grapholitha molesta, Helicoverpa spp. , Leucoptera seitella, Lithocolletis blaneardella, Lobesia botrana, Loxostege stictie a- Iis1 Disparate Lymantria, Lymantria monaeha, Lyonetia elerkella, Manduca Friday, Malacosoma neustria, Mamestra brassicae, Mocis repanda, Operophthera brumata, Orgyia pseudotsugata, Ostrinia nubilalis, Pandemis heparana, Panolis flammea, Pectinophus citrus, Phylliophus operus Iheris brassicae, Iheris rapae, Plathypeena scabra, Platynota stultana, Plutella xylostella, Prays citri, Prays oleae, Iphenenia sunia. , Spodoptera littoralis, Spodoptera litura, Syllepta derogata, Synanthedon myopaeformis, Thaumatopoea pityo- eampa, Tortrix viridana, Trichoplusia ni, Tryporyza ineertulas, Zeiraphera eanadensis; especially Heliothis spp., Helicoverpa Spp., Crocidolomia pavonana, Pieris rapae, Phthorimaea operculella, Chrysodexis Spp., and Plutella xylostella;

(b) of the order Hemiptera (Hemiptera), for example Aphis, Bemisia, Phorodon, Aeneolamia, Empoasca, Parkinsiella, Pyrilla, Aonidiella, Coccus, Pseudococcus, Helopeltis, Lygus, Dysdercus, Oxycarenus, Neza, Aleyrodes, Aleyrodes, Aleyrodes Triatoma, Psylla, Myzus, Megoura, Phylloxera, Adelges, Nilaparvata, Nephotettix or Cimwx spp .;

(c) of the order Orthoptera (Orthoptera), for example Gryllotalpa gryllotalpa, Migratory locust, Melanoplus bivittatus, Melanoplus femur rubrum, Melanoplus mexicanus, Melanoplus spretus, Nomadacris septemfasciata, Schistocerus americanus, Schistocerus americanus , Schistocerca gregaria;

(d) of the order of psocoptera (Psocoptera), for example, Peripsocus spp .;

(e) of the order of the hymenoptera (IIymenoptera), for example, Athalia rosae, Atta cephalotes, Atta sexdens, Atta texdens, Hoplocampa minuta, Hoplocampa testudinea, Iridomyrmex purpureus, Monomorium pharaonis, Solenopotes eap Solopsisen, Solenopotes eap Solenopsis richteri, Technomyrmex albipes;

(i) of the order of termites (Isoptera), for example, Calotermes flavicolis, Coptotermes spp, Leucotermes flavipes, Macrotermes subhyalinus, Nasutitermes spp, such as Nasutitermes walkeri, Odontotermes formosartus, Reticulitermes lucifugus, Termes natalensis;

(g) beetles (Coleoptera), for example, Anthonomus grandis, Anthonomus pomorum, Apion vorax, Atomaria linearis,

Phagus piniperda, Cassida nebulosa, Cerotoma trifurcata, Ceuthorhynchus assimilis, Ceuthorhynchus napi, Chaetocnema tihialis, Conoderus vespertinus, Cryoceris asparagi, Dendroctonus refipennis, Diahrotica longicomis, Diahrotis viriphis, Ephrothis varifera abie- tis, Hypera brunneipennis, False hypera, Ips typographus, Lemma bilineata, Lemma rnelanopus, Leptinotarsa decemlineata, Limonius californicus, Lissorhoptrus oryzophilus, Meligothes aeneus, Melolontha hiprushachus, Orphomachus oculus , Phaedon eochleariae, Phyllopertha horticola, Phyllophaga sp., Phyllotreta ehrysoeephala, Phyllotreta nemorum, Phyllotreta striolata, Popillia japoniea, Psylliodes napi, Scolytus intrieatus, Sitona lineatus, Sitophilus granarius;

(h) of the order Dictyoptera, for example, from families Polyphagidae, Bladberidae, Blattidae, Epilampridae, Chaeteesidae, Metallyeidae,

Mantoididae, Amorphoscelid.ae, Eremiaphilidae, IIymenopudidae, IvIantidae and Empusidae;

(i) of the order of tripods (Thysanoptera), for example, Frankliniella fusca, Frankliniella occidentalis, Frankliniella tritici, Haplothrips tritici, Heliothrips haemorrhoidalis, Scirtothrips citri, Thrips oryzae, Thrips.

palmi, Thrips tabaci;

(j) of the order of the homoptera (Homoptera), for example, Acyrthosophon onobrychis, Acyrthosiphon pisum, Adelges laricis, Aonidiella aurantii, Aphidula nasturtii, Aphis fabae, Aphis gossypii, Aphis pomi, Aulacorisia, Brachycaudus cardui, Brevicoryne brassica- and Dalbulus maidis, Dreyfusia nordmannianae, Dreyfusia piceae, Dysaphis radicola, Empoasca fabae, Eriosoma lanigerum, Laodelphax striatel-la, Macrosiphum avenae, Macrosiphum euphoriae, myrosphorae myeophorbiae , Myzus cerasi, Nephotettix cincticeps, Nilaparvata lugens, Perkinsiella saccharicida, Phorodon humuli, Psylla mali, Psylla piri, Psylla pyricola, Rhopalosiphum maidis, Schizaphis graminum, Sitobion avenae, Sogatella furcifleida vaporis, Togonotria vitrius,

(k) Diptera (Diptera), for example Anastrepha ludens, Ceratitis capitata, Contarinia sorghicola, Dacus cucurbitae, Dacnxs oleae, Dasineura brassicae, Delia radicum, Delia radicum, Hydrellia griseola, Hylemyia platura, Liriomyza trifoliae, Liriomyza trifoliae Lucilia Sp., Mayetiola destructor, Musca sp., Orseolia oryzae, Oscinella frit, Pegomya hyoscyami, Phorbia antiqua, Phorbia brassicae, Phorbia coarctata, Khagoletis cerasi, Rhagoletis pomonella \

(1) of the order Phthiraptera (Anoplura), eg Pthirus pubis, Pediculus humanus capitus, Pediculus humanus humanus, long-nosed sucking louse, linognathus vituli, short-nosed sucking louse, IIaematopinus eurysternus, small blue louse, Solenopotes capillatus, buffalo louse Haematopinus tuberculatus, and tail switch louse Haematopinus quadripertusis and Mallophaga, for example of the genus Bovicola, such as Bovicola ovis or Bovicola bovis, Durnulania, Trichodectus and Menopon; especially Bovicola ovis or Bovicola bovis; (m) of the order of the siphonaptera (Siphonaptera), for example Ctenocephalides or Pulex spp.

(n) of the order Blattodea, including Periplaneta Americana, Blattella germanica and Blattela asahinai;

(o) Dermaptera, which are the earwigs;

(p) of the order Arachnida, for example, Ixodes holocyclus, Boophilus microplus, Rhipicephalus sanguineus, Sarcoptes seabiei var. humani, Sarcoptes seabiei canis, Sarcoptes seabiei suis, Sarcoptes seabiei bovis, Psoroptes ovis and Dermatophagoides spp., especially Sarcoptes seabiei var. humani, Sarcoptes seabiei canis, Sarcoptes seabiei suis, Sarcoptes seabiei bovis, Psoroptes ovis and Dermatophagoides spp. Especially preferred plant-infesting pests include IIelicoverpa spp., Such as Helicoverpa armigera, Helieoverpa Zea and Helieoverpa punetigera (Budworms), Pavonana Croeidolomia (Cabbage Head Caterpillar), Phens rapae (White Cabbage Butterfly), Phthorriariaea operculella ( potato moth), Chrsyodexis spp. (Tobacco loopers), Plutella xylostella (Crucifix moth) and Epiphyas postvittana (Walker) (apple moth), Cydia pomonella (Apple moth), Weevil spp, Aphelenchoides (leaf nematodes), Meloidogyrie (gall nematodes) Heterodera, Globodera (cyst nematodes), such as potato nematodes, Nacobhus, Pratylenehus (lesion nematodes), Ditylenehus, Xiphinema, Longidorus, Trichodorus Meloidogyne and Xiphine index.

Especially preferred pests infesting domestic animals include Bovieola ovis (sheep louse), Bovicola hovis, Haemato-pinus eurysternus (short-nosed cattle louse), Linognathus vituli (long-nosed cattle louse), Solenopotes eapillatus (tubereule -hearing lousé), Sareoptes seabiei canis (scabies), Sareoptes seabiei suis, Sareoptes seabiei bovis, Psoroptes ovis, Haemonehus eontortus, II. placei, Ternchostrongylus eolubriformis, Ternchostrongylus eolubriformis, and Cooperia spp.

Especially preferred pests infesting humans include

in IyIhirus pubis, Pediculus humanus capitus, Pediculus humanus humanus, Sareoptes seabiei var. humani and Dermatophgoides spp.

In one embodiment, the pest which is prevented from undergoing a remodeling event by the present invention is selected from the group consisting of louse, flea, tick, fly, mite and other mincing or blood-sucking pests. In one embodiment, the pest egg is a louse egg, more preferably a head louse egg. Lice are parasites that feed on animal skin and blood and they deposit their digestive juices and fecal material into the skin. These materials, as well as the sting wound, cause irritation and resulting scratching skin lesions, and can cause a serious infection with ganglion inflammation. Head lice are also vectors of certain diseases, such as epidemic or exanthematic typhus and recurrent fever. The adult female louse has a life span of about one month and can lay up to 10 eggs per day. Lice that may infect humans may include the species of crab lice (L ^ thirus pubis) and the separate species Pediculus humanus, which is composed of two subspecies, Pedicnxlus humanus capitis or Pediculus humanus humanus or body louse (Busvine). , Antenna, 1993, 17: 196-201). The above subspecies of lice are closely related and are known to successfully crossbreed each other in the laboratory situation (Busvine, Cuta- neous Infestations and Insect Bites, 1985, 163-174).

The head louse Pediculus humanus var. capitis, is a specific host ectoparasite that lives exclusively on human heads and feeds on scalp blood suction. Following a blood meal, the mature adult female louse will lay up to 10 eggs near the scalp for a period of 24 hours. The eggs are firmly attached to the hair shaft by a glue. Seven to ten days after laying, depending on temperature and humidity, eggs will hatch and emerging nymphs will begin to feed. The nymphs progress through three seedlings (1st instar, 2nd instar, 3rd instar), with each seedling taking 3 to 5 weeks to complete. Following the final moult, the adult male or female emerges, with mating occurring two days later. Within hours of feeding, eggs will be produced and the cycle continues. The entire life cycle from egg to egg takes approximately 20 to 30 days to complete, depending on the heating and humidity conditions. Following hatching of the egg, the eggshells remain attached to the hair shaft and will gradually move away from the scalp as the hair grows. Hatched eggs (nits) are relatively easy to detect due to their refractive nature, appearing white under artificial light, in contrast, un hatched eggs have a slightly pale brown color, allowing them to match most hair colors and thus making it difficult to detect.

In another embodiment, the pest that is prevented from undergoing a remodeling event by the present invention is one that infests a plant host, including plant roots. In a preferred embodiment, the pest is a budivorm egg, a caterpillar egg, a butterfly, a moth or a root nematode. Caterpillars, butterflies, moths, soil nematodes and their larvae feed on valuable crop plants, * 10 such as cotton, oilseed crops such as canola, ornamentals, flowers, fruit trees, cereal crops, vines, tuber crops, grassland, tobacco, legumes and vegetables, especially kale crops such as cauliflower and broccoli, cotton, maize, sweet corn, tomatoes, tobacco and legumes such as soybeans, beans , guinea beans, guandu and chickpeas.

Cruciferous moth larvae (Plutella xylostella) feed on all plants of the Brassica / cruciferae family, including canola and mustard, plant crops such as broccoli, cauliflower and cabbage, and also on various greenhouse plants. Usually, the cruciferous moth takes about 32 days to develop from egg to adult. However, depending on food and weather conditions, a generation may take 21 to 51 days to complete. Adult female moths lay an average of 160 eggs over a life span of about 16 days. A female will lay eggs at night and lay the largest number of eggs on the first night after emerging from the pupa. The eggs are small, spherical or oval and whitish yellow and are stuck to the top and bottom surfaces of a leaf either alone or in groups of two or three. Eggs are usually laid along leaf veins where the leaf surface is uneven. The eggs hatch in about five to six days. After hatching, the larvae burrow into the leaf and begin to eat the leaf tissues internally. After about a week, the larvae come off the leaf and feed externally. The larvae undergo molting three times for 10 to 21 days and at maturity they are about 12 mm long. The larvae make pupae in delicate open-mesh cocoons attached to the leaves and the pupa stage lasts for 15 days.

Budworrns, such as corn cob caterpillar, tomato bug, cotton apple caterpillar, and cotton bollworm are serious pests in a number of crops, such as sunflowers, zucchini, beans, * 10 peppers, alfalfa, potatoes, leeks, cotton, maize, plums, citrus plants, tomatoes, tobacco and lettuce, and flowers such as geraniums and roses. Budworm occurs in many regions of the world and in temperate climates may have 2-3 generations per season with hibernation in pupae in the soil. In tropical regions, budworrns can continue to be active during the year. Eggs are small (~ 0.5 mm in diameter) and dome-shaped with a slightly flat bottom. Eggs are usually laid singularly next to buds or parts of flowers or leaves. An adult can lay 500-3000 eggs. Eggs hatch after only three days at 25 ° C or longer at cooler temperatures, eg 9 days at 17 ° C. The larval feeding period is about 19 to 26 days under favorable temperature and feeding conditions and when fully developed, the larvae move to the ground to make pupa. The pupal period usually lasts from 8 to 21 days, although diapusal pupae may hibernate in temperate regions.

In another embodiment, the pest that is prevented from

The remodeling method of the present invention is one that internally infests a human or animal. In a preferred embodiment, the pest is a nematode, trematode or cestoid. Nematodes (lower backs), trematodes and cestodes are flat worms and can cause significant damage to humans or agriculturally important animals such as sheep, cows, pigs and goats.

Haernonchus contortus, an intestinal parasite that infests sheep and goats, and adult male and female worms live in the abomasum or the true stomach of ruminant animals. Female worms lay 5,000 to 10,000 eggs per day, which leave the host with the feces. After hatching, young people in the first and second stages feed on bacteria. Youngsters in the third stage keep a cuticle like a sheath and the youngsters in the third stage are ingested by the host while grazing. Young sheathed worms pass * 10 to the host and shed before entering the abomasum. In abomasum the spilled young worms burrow in the mucosa and feed on blood. Once adulthood is reached, mating occurs and other eggs are laid. The entire life cycle of this parasite takes approximately 21 days. Infested sheep may suffer Ul thrift resulting in weight loss and large infestations may result in anemia, which, left untreated, may cause the animal to die.

In one embodiment of the present invention, the methods and compositions are for treating or preventing external infestation of a human or animal by a pest or parasite undergoing remodeling events such as lice, fleas, mites or ticks by inhibiting such events. remodeling. Inhibiting remodeling events has the advantage of disrupting pest or parasite life cycles and / or reproduction cycles thereby controlling the infestation.

In another embodiment the methods and compositions are for treating or preventing internal infestation of a human or animal by a pest or parasite undergoing remodeling events, such as nematodes and trematodes, by inhibiting the transition from one stage of the pest or parasite life cycle. for the next. Inhibiting remodeling events has the advantage of disrupting pest or parasite life cycles and reproduction cycles at a number of different points thereby controlling the infestation.

In yet another alternative embodiment, the methods and compositions are for treating or preventing infestation of an environment with a pest or parasite by inhibiting pest or parasite remodeling events.

For example, pest or parasite eggs may be laid

on soil around a plant, on carpet or curtains in a house (eg, flea eggs), bedding or mattresses (eg, mite eggs or bedbug eggs) or on or near structures such as buildings or other wood products (for example, termite eggs). The hatching of eggs allows reinfestation of humans, animals or plants in the environment or damage to products in the environment. Inhibiting remodeling events has the advantage of disrupting pest or parasite reproduction cycles, thereby controlling infestation. In addition, preventing reinfestation results in a reduction in the number of pesticide applications required to control an infestation.

In yet another alternative embodiment of the present invention, the methods and compositions of the invention are for treating or preventing larval infestation of plants by inhibiting remodeling events. The present applicants have identified metal chelating agents as effective agents to inhibit remodeling events affecting both eggs and larvae that feed on commercially valuable plants. The use of metal chelating agents to inhibit remodeling events has the advantage of inhibiting reproduction cycles of commercially valuable plant-feeding larvae, thereby controlling the pest infestation of commercially valuable plants.

The term "metalloprotease" as used herein is used to refer to a protease involved in invertebrate remodeling events during one or more stages of development of a pest species, wherein the protease has an active metal ion that acts as a catalyst. Preferably, the metalloprotease contains a zinc ion that participates in catalysis by polarizing a water molecule to attack a peptide substrate bond. More preferably, metalloprotease is sensitive to metal chelating agents that are capable of directly or indirectly blocking its activity. Metalloprotease may be involved in egg hatching induction by acting on an egg's operculum to facilitate egg hatching, or it may reduce the strength of the egg shell, allowing the nymph or lavas to dislodge the shell during hatching. Metalloprotease may also be involved in facilitating the change from a larval or immature stage to a subsequent stage and also to mature or adult form. Metalloprotease may be directly or indirectly involved in remodeling events. Suitable metalloproteases involved in remodeling events may include endoproteases (enzymes that break down within the peptide chain) and exoproteases (enzymes that break down amino acid (s) from peptide termini). Exoproteases may further be categorized as carboxiproteases (which break down C-terminal amino acid (s)) or aminopeptidase (which break down N-terrinal amino acids). MctalocarboxypioLeases require a bivalent cation (usually Zn2 +) for activity, while aminopeptidases are generally classified according to their dependence on metal ions (Zn2 + or Mg2 +). They exist in both free and membrane bound forms and high pll assist activity (8-10). One method of detecting metalproteases associated with egg hatching may involve collecting the surrounding fluid from the developing embryo both at the time of egg hatching and washing empty eggshells shortly after hatching and analyzing the sample for the presence of eggs. proteases using SDS-PAGE analysis with gelatin substrate. Having shown the presence of proteolytic activity of the sample it is then possible to incubate the sample in the presence of a metalloprotease inhibitor which has been identified as having the required arrangement of polar atoms and clogP values and / or molar refractivity and in some embodiments a preference. by zinc ion chelators, and then reanalyze the treated samples to determine whether the activity of proteases extracted from the egg was inhibited. Having shown inhibition of metalloprotease activity (s) obtained from hatched eggs, it is then possible to expose un hatched eggs, for example, to the same inhibitor and to evaluate whether inhibition of egg hatching occurs. A similar approach can be taken to determine appropriate metal chelating agents to inhibit other remodeling events such as pololysis, ecdysis, shedding or metamorphosis. For example, fluid may be obtained from invertebrates undergoing polysis, ecdysis or metamorphosis and the presence of proteases detected as described above. Suitable metal chelating agents can then be determined. Metalloproteases involved in egg hatching can also be identified by identifying a gene encoding a metalloprotease, silencing that gene and showing that the egg is unable to hatch by methods known to those skilled in the art.

The phrase "inhibiting remodeling events" as used in this document is used to mean inhibition of protease enzymes involved in remodeling events involving the envelopes of invertebrate multicellular organisms, eg eggs, sheaths, shells, exoskeletons. , cysts, cocoons or ootheca. In the present invention, a particular life cycle stage of the invertebrate pest is exposed to a metal chelating agent that is capable of preventing a remodeling event when compared to the same untreated life cycle stage. In the case of egg hatching, this remodeling event may be characterized by hatchflap or operculum of the hatched egg and shortly thereafter the emergence of a larva or nymph. In the case of the louse, the head appears first, followed by the chest to which the legs are attached. Finally, the abdomen emerges and the nymph gets rid of the egg. In the case of a moth or butterfly egg, the eggshell is weakened by the action of protease enzymes and the emerging larva breaks the eggshell. Egg hatching is taken to exclude damage or accidental rupture of an eggshell.

Preferably, the metal chelating agent is a compound capable of inhibiting remodeling events when it is applied to a stage of the pest life cycle at any time between placement and during adulthood.

The remodeling event preferably occurs in a pest present in, but not limited to, a host organism, such as the skin, fur, fur or fleece of an animal or skin or fur, such as hair of a human. In alternative embodiments of the invention the remodeling event occurs in a pest present in host plants or in plant roots including cereal crops, fruit trees, cotton, oilseed crops, ornamental plants, flowers, vine crops, tuber crops. , pasture and vegetable crops. In yet other alternative embodiments, the remodeling event occurs in a pest that is present in a breeding environment or place, such as, but not limited to, houses and buildings, pet or farm enclosures, carpets, bedding such as sheets and blankets, curtains and furniture. In yet other alternative embodiments, the remodeling event occurs in a pest that is within a host, such as, but not limited to, humans, farm and domestic animals.

In accordance with the present invention, the pest may be exposed to a metal chelating agent by any appropriate means. One skilled in the art will appreciate that these media may vary widely, depending on whether the chelating agent should be applied to a host, such as a plant, or applied to or administered to an animal, including a human, or applied to various environments in others. breeding sites, and depending on the nature and type of the targeted pest. Suitable means for exposing the pest present in animals to metal chelating agents include, but are not limited to, direct topical application, such as by dipping or spraying, implanting, delayed release formulations or devices, or orally. Where the invention is applied to humans, formulations suitable for topical application include, but are not limited to, sprays, aerosols, shampoos, musses, creams and lotions, and formulations suitable for internal application include, but are not limited to, tablets, capsules or formulations. net. In some situations, parenteral injection may be the most appropriate means of treatment for humans or animals. Where metal chelating agent is to be applied to plants, suitable means include, but are not limited to, sprays, powders, including wettable talc, wettable granules and suspension concentrates, lozenges, liquids including microencapsulation and aerosols. The method of the invention also encompasses the concurrent or successive use of two or more metal chelating agents or the use of one or more metal chelating agents in conjunction with or in succession with other known pest controlling agents.

In yet another aspect of the invention, the methods and compositions may include other hatching pesticides, larvae, nymphs or adult pests. For example, suitable pesticides which may be used in conjunction, either simultaneously, separately or sequentially with the metal chelating agents of the present invention include macrocyclic lactones such as spinosad, botanical insecticides, carbamate insecticides, dehydrating insecticides, dintrophenol insecticides, fluoride insecticides, formamidine insecticides such as armitraz, fumigant insecticides, inorganic insecticides, insect growth regulators (including chitin synthesis inhibitors, juvenile hormone analogs, juvenile hormones, mute hormone antagonists, mute hormone antagonists, molting hormones, molting inhibitors), nicotinoid insecticides, organochlorine insecticides, organophosphate insecticides, heterocyclic organothiophosphate insecticides, phenyl organothiophosphate insecticides, phosphonothioate insecticides, phosphoramide phosphate insecticides, midothiatos, phosphorodiamide insecticides, oxadiazine insecticides, phthalimide insecticides, pyrazole insecticides, pyrethroid insecticides, pyrimidinamine insecticides, pyronic insecticides, thiourea insecticides and urea insecticides, including agents described in EP 0191236, US 5,288,483 and US 6,727,228. Other useful insecticides include dimethicone copolyols, such as those described in US 6,663,876 and US 6,607,716, which have low toxicity. Useful nemiticides that may be used include Oxfendazole, Albendazole, Mebendazole / Closantel, Fenbendazole and Triclabenzazole. Trematode and cestode infection can also be treated with Praziquantel.

The metal chelating agent may be applied to a host's hair or skin when the host is a human or animal, preferably in a region that is infested with a pest. The infestation may be due to pests selected from the group consisting of lice, fleas, ticks, mites and other biting or blood-sucking pests, and combinations thereof. The metal chelating agent may be applied topically in the form of ointments, aqueous compositions including solutions and suspensions, creams, lotions, aerosols, sprays or talc. When the pest infests the human or animal internally, the metal chelating agent may be applied or administered internally, for example in the form of a tablet, capsule or ingestible liquid formulation. When the host is a plant, the pest infestation is preferably due to selected pests of larvae, butterflies, pests or nematodes. The metal chelating agent may be applied topically, for example as a spray or powder. When the infestation is in the environment, such as a termite infestation, the metal chelating agent may be applied in a formulation such as a spray, fumigant or powder.

The term "effective amount" means a concentration of at least one metal chelating agent sufficient to provide treatment or prevention of a pest infestation in a host or environment. The effective amount of a metal chelating agent used in the methods of the present invention may vary depending on the host and the type and level of infestation. In one embodiment, the metal chelating agent is applied to the scalp of a person suffering from head louse infestation and is left on the treated person for a period of time to prevent hatching of louse eggs. Preferably, the time period is between 5 and 15 minutes. The metal chelating agent is preferably used at a concentration of from about 0.100mM to 1M, preferably 0.01mM to 100mM, more preferably in the range 0.1mM to 100mM. The effective amount depends on the metal chelating agent used. However, some dipyridyl components may conveniently be applied in the range of 5mM to 100mM, especially at a level of about 50mM. Appropriate amounts of compounds of formula (II), such as tetralone compounds, may be applied at a level in the range of 0.5 mM to 100 mM, especially 50 mM ImM. Since a significant number of mammalian proteases require zinc for their activity and can be affected by metal chelating agents, it would be necessary to ensure that the metal chelating agent has been used in a safe and effective amount and preferably targets especially in a remodeling event. specific, such as egg hatching, apolysis, ecdysis, shedding or metamorphosis.

In another embodiment the metal chelating agent is applied to a commercially valuable plant to prevent remodeling events occurring in a pest that are involved, for example, in egg hatching or moulting. The chelating agent can be applied directly or indirectly to pests that are present in the soil or leaves, buds, stems, flowers or fruits of a plant by spraying, brushing or dusting. Suitable compositions include emulsifiable concentrates, directly dilutable or sprayable solutions, dilute emulsions, wettable talc, talc, powder or soluble granules. The metal chelating agent is preferably used at a concentration of from about 0.0001mM to 1M, preferably 0.01mM and 100mM, more preferably in the range of 0.1mM and 30mM. The effective amount depends on the metal chelating agent used. However, some dipyridyl compounds may suitably be applied in the range of 5 mM to 15 mM, especially at a level of about 10 mM.

Appropriate amounts of compounds of formula (II) include, but are not limited to, the range from 10 mM to 20 mM, especially 10 mM to mM.

The host treated by the methods of the invention may be selected from, but without limitation, the group consisting of humans, sheep, cattle, horses, pigs, poultry, dogs and cats. The methods of treatment or prevention of the present invention may be applied to plants and or other pest breeding sites. Plants or their roots treated by the methods of the invention are preferably selected from the group consisting of cotton, oilseed crops such as canola, ornamentals such as shrubs, flowers such as chrysanthemum, michaelmas daisy, geranium and roses, fruit trees such as apple, pear, plum, kiwi, gooseberry and citrus varieties, for example lemons, oranges, limes and grapefruits, cereals such as maize and sweet corn, vine growing, such as grapes, tubers, grazing plants such as meadows and white clover, lucerne and lupins, and vegetables such as cabbage, for example broccoli and cauliflower, cabbage, tomatoes, zucchini, garlic -pork, lettuce, and beans, as well as legumes such as beans, soybeans, guandu, chickpeas, and chickpeas.

The compositions of the present invention may be formulated as solutions and emulsions. Suitable excipients such as emulsifiers, surfactants, stabilizers, dyes, penetration enhancers and antioxidants may also be present in the compositions. Suitable carriers which may be added to the compositions may include water, saline, alcohols, polyethylene glycols, gelatin, lactose, magnesium stearate and silicic acid. The compositions may include sterile and non-sterile aqueous solutions. In one embodiment, the compositions are in a soluble form and the metal chelating agent is diluted in a soluble sterile buffered saline or water solution. The compositions may also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions may further contain substances that increase the viscosity of the suspension and may also contain stabilizers. The solutions may also contain buffers, diluents and other suitable additives. The compositions may go alongside other adjuncts than they are.

X 5 JT - i ~ ------- - Ji

compatible with metal chelating agent activity. The compositions of the present invention may be formulated and used as foams, emulsions, microemulsions, shampoos, musses, creams and jellies. The formulations of the compositions described above would be known to those skilled in the field of pesticides.

The active ingredients according to the invention may be used

to inhibit remodeling events that occur in plant pests or their roots, especially in crops of useful and ornamental plants in agriculture, horticulture and forestry, or in parts of such plants such as fruits, flowers, foliage, stems , tuber or root, and in some cases even parts of plants that are formed at a later time, protection against these pests is granted. In these compositions, the active ingredient is employed together with at least one of the auxiliaries conventionally used in the formulation art, such as extenders, for example, solvents or solid carriers, or such as surface active compounds (surfactants).

Examples of suitable solvents are partially hydrogenated or unhydrogenated aromatic hydrocarbons, preferably C8 -C12 alkylbenzene moieties, such as mixtures of xylene, alkylated or tetrahydronaphthalene, aliphatic or cycloaliphatic hydrocarbons, such as paraffins or cyclohexane, such as methanol, ethanol, propanol or butanol, glycols and their ethers and esters, such as propylene glycol, dipropylene glycol ether, hexylene glycol, ethylene glycol, diethylene glycol, ethylene glycol monomethyl ether or ethylene glycol monoethyl ether, ketones, such as such as cyclohexanone, isophorone or diacetone alcohol, strongly polar solvents such as N-methylpyrrolidine-2-one, N-methylpyrrolidine, dimethyl sulfoxide or N, N-dimethylformamide, water, free or epoxidized rapeseed, castor oil, coconut or soy, and silicone oils.

Solid carriers which are used, for example, for dispersible powders and talc are, as a rule, natural minerals from the ground such as calcite, mica, kaolin, montmorillonite or attapulgite. To improve physical properties, it is also possible to add highly dispersed silicas or highly absorbent polymers. Particle adsorptive carriers suitable for granules are porous types such as pumice, brick dust, sepiolite or bentonite, and suitable non-sorting carrier materials are calcite or sand. In addition, a large number of granular materials of organic or inorganic nature may be used, in particular dolimite or pulverized plant residues. Suitable surface active compounds are, depending on the nature of the active ingredient to be formulated, nonionic, cationic and / or anionic surfactants or mixtures of surfactants that have good emulsifying, dispersing and wetting properties. The surfactants listed below are for examples only; Many more surfactants conventionally used in the formulation art and suitable in accordance with the invention are described in the relevant literature.

Suitable nonionic surfactants are primarily derived from polyglycol ether of aliphatic or cycloaliphatic alcohols, saturated or unsaturated fatty acids and alkylphenols, which may contain from 3 to 30 glycol ether groups and from 8 to 20 carbon atoms in the hydrocarbon radical (aliphatic ) and from 6 to 18 carbon atoms in the alkyl radical of alkylphenols. Water soluble polyethylene oxide adducts with polypropylene glycol, ethylene diamine polypropylene glycol and alkyl polypropylene glycol having from 1 to 10 carbons in the alkyl chain and from 20 to 250 of ethylene glycol ether and from 10 to 100 propylene groups are also suitable. glycol ether. The above-mentioned compounds usually contain from 1 to 5 units of ethylene glycol per propylene glycol unit. Examples that may be mentioned are nonylphenylpolyethoxyethanols, castor oil polyglycol ethers, polypropylene / polyethylene oxide adducts, tributylphenoxypolyethoxyethanol, polyethylene glycol and octylphenoxypolyethoxyethanol. Polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene sorbite trioleate are also suitable.

Cationic surfactants are primarily quartenary ammonium salts, which have at least one alkyl radical of 8 to 22 carbon atoms and, as other substituents, lower alkyl, benzyl or lower hydroxyalkyl radicals which may be halogenated. The salts are preferably in the form of halides, methylsulfates or ethylsulfates. Examples are stearyltrimethyl ammonium chloride and benzyldi (2-chloroethyl) ethylammonium bromide.

Suitable anionic surfactants may be both water soluble soaps and water soluble synthetic surface active compounds. Soaps which are suitable are alkali metal salts, alkaline earth metal salts and substituted or unsubstituted ammonium salts of higher (C10-C22) fatty acids, such as sodium or potassium salts of oleic or stearic acid, or mixtures of natural fatty acids, which may be obtained, for example, from coconut oil or liquid pitch; or fatty acid methyltaurinates. However, synthetic surfactants, in particular fatty sulphonates, fatty sulphates, sulphonated benzimidazole derivatives or alkylaryl sulphonates, are most often used. As a rule, fatty sulfonates and fatty sulfates exist as alkali metal salts, alkaline earth metal salts or substituted or unsubstituted ammonium salts and generally have an alkyl radical of 8 to 22 carbon atoms, alkyl also including the following alkyl of acyl radicals. Examples of fatty sulfonates and fatty sulfates include the sodium or calcium salt of lignosulfonic acid, dodecylsulfuric ester or a fatty alcohol sulfate mixture prepared with natural fatty acids. This group also includes the salts of sulfuric and sulfonic acid esters of ethylene oxide / fatty alcohol adducts. The sulfonated benzimodazole derivatives preferably contain 2 sulfo groups and a fatty acid radical having from about 8 to 22 carbon atoms. Examples of alkylarylsulfonates are the sodium, calcium or triethanolammonium salts of dodecylbenzenesulfonic acid, dibutyl naphthalenesulfonic acid or a formaldehyde naphthalenesulfonic acid / condensate. Corresponding phosphates such as salts of the phosphoric ester of a p-nonylphenol (4-14) ethylene oxide adduct or phospholipids are also suitable. In a preferred embodiment, the composition comprises a metal chelating agent at a concentration of about 100,000 to 1M, preferably between 0.1mM to 100mM, more preferably in the range of 50mM to 0.1mM. Compositions containing some metal chelating agents, for example the compounds of formula I, may preferably contain between 5 and 50mM of the compound, especially at a level of about 30mM. Compositions containing compounds of formula (II) may preferably contain between 0.1mM to 100mM, especially 50mM to 10mM.

A compound that inhibits egg hatch remodeling events in a pest may be identified using a method comprising assessing the clogP value and / or molar refractivity of the compound and / or the ability of the compound to bind zinc and / or inhibit an involved metalloprotease. at the remodeling event.

In another aspect of the invention, there is provided a method of selecting a chelating agent as a candidate inhibitor of invertebrate remodeling events from a collection of metal chelating agents, said method comprising selecting metal chelating agents having at least two polar atoms. able to simultaneously coordinate with a metal ion and

(i) a clogP value of / 1 and <4; and / or

(ii) a molar refractivity in the range of 40 to 90 cm3 / mole;

The clogP of a metal chelating agent can be calculated from its IogP value using a clogP program, for example, the program provided by Biobythe. IogP values can be obtained from the literature or can be calculated from a measured partition coefficient between n-octanol and water.

The molar refractivity of a metal chelating agent can be calculated using Biobythe's CMR (calculated molar refractivity) computer program.

In some embodiments, the metal chelating agent is selected to inhibit a zinc-dependent metalloprotease enzyme involved in an invertebrate remodeling event. In such cases, the chelating agent to bind zinc ions can be determined by determining the constant association (IogKb) of the zinc metal chelating agents. Constant association can be determined from the literature using methods known to those skilled in the art. In preferred embodiments in which the metalloprotease enzyme is a zinc-dependent metalloprotease enzyme, metal chelating agents having a constant zinc association of more than 5.0 are selected.

A similar procedure may be followed if the metalloprotease to be inhibited includes a metal ion other than zinc, for example Mg ++, Cu ++ or Fe ++. The constant association of the metal chelating agent to that metal ion can be evaluated and the metal chelating agents having the highest association constants can be selected.

Identification of suitable metal chelating agents may further comprise testing the compound in a biological assay. A suitable biological assay preferably comprises exposing a pest control sample in which the remodeling event may occur to a control buffer or control formulation while at the same time exposing a pest test sample in which remodeling may also occur to a remodeling event. solution or formulation comprising a test compound.

A compound that is effective in inhibiting a remodeling event in a pest is identified when the remodeling event is observed in the sample pests or control formulation and the remodeling event is not observed in the pest test sample. In the biological assay of the present invention, the remodeling event may occur in a pest selected from the group consisting of lice, fleas, ticks, flies, mites and other biting or blood-sucking pests and further includes pests that live within a host. mammals such as trematodes, nematodes and cestodes. In the biological assay of the present invention, the remodeling event may occur in plant infesting pests such as caterpillars, moths, butterflies and soil nematodes. Alternatively, the remodeling event occurs in a pest that infests an environment, such as a termite egg or mite egg.

The control buffer may include, but is not limited to, sterile phosphate buffered saline or water or an organic solvent. The compound tested is preferably a metal chelating agent. In one example of a biological egg hatching assay, egg hatching is observed when the hatchflap or egg operculum opens and then larvae or nymphs begin to emerge. In the case of head lice, the head appears first, followed by the chest to which the legs are attached. Finally, the abdomen comes out and the nymph gets rid of the egg. In the case of head lice, the eggshells then remain attached to the hair shaft. A metal chelating agent test compound may be identified as suitable for use in the invention if the eggs exposed to the control buffer exhibit a high hatching level, for example 70-100%, whereas the egg exposed to the control chelating agent. Test metal exhibits a low level, eg 0-30%, of egg hatching, especially where 100% of egg hatching inhibition occurs.

In preferred embodiments, the metal chelating agent pLDso is greater than 2, preferably greater than 3, and more especially greater than 4.

Other biological assays can be used to evaluate the activity of selected metal chelating agents in inhibiting other remodeling events, such as encystation, shedding, apolysis, ecdysis or metamorphosis. For example, an invertebrate at a particular stage of life, for example, a cyst, larva, cocoon, pupa, nymph, or adult, may be exposed to a test metal chelating agent in a vehicle and the occurrence of a remodeling event, such as encystment, shedding, apolysis, ecdysis or metamorphosis observed and compared with a vehicle-exposed invertebrate control group in the absence of test metal chelating agent.

In another aspect of the invention there is provided a use of at least a metal chelating agent in the manufacture of a composition for inhibiting a protease enzyme involved in invertebrate remodeling events or for treating or preventing pest infestation, wherein at least least one chelating agent has at least two polar atoms capable of simultaneously coordinating with a metal ion and

(i) a clogP value of / 1 and <4; and / or

(ii) a molar refractivity in the range of 40 to 90 cm3 / mole;

or a pharmaceutically acceptable, acceptable or agriculturally acceptable salt thereof.

In one embodiment, the pest is one that infests a host plant. In another embodiment, the plague is one that infests a domestic animal. In yet another alternative embodiment, the pest is one that infests an environment.

Agents comprising at least one metal chelating agent are also encompassed by the present invention, as described herein, to inhibit a protease enzyme involved in invertebrate remodeling events or to treat or prevent pest infestation. In some aspects of the invention it will be possible to identify additional pesticidal agents for inhibiting invertebrate infestation of a host. As noted above, it has been found that compounds of Formula I and II are particularly useful in inhibiting various invertebrate remodeling processes and / or invertebrate metabolic processes. The rational drug design strategy can be used to specifically identify these inhibitors. It is now established in the present invention that compounds of Formula I and II have properties useful in inhibiting invertebrate proteases. Drug design strategies can be created in which each of the R groups in the Formula I and Formula II core structure is separately and individually fixed and the effectiveness of the resulting agent in a given assay is determined. The structure of Formula I and Formula II that specifically interacts with, for example, protease can be modeled using computational tools. These tools can allow a drug molecule to be constructed within the biomolecule using knowledge of its structure and the nature of its active site.

Compounds tested for efficacy as pesticides may be part of a compound set or library, which may be a diverse set or library or a centered set or library, as will be apparent to the skilled person. Libraries that may be used for such screening may be prepared using combinatorial chemical processes known in the art or conventional means for chemical synthesis. Collections of compounds of formula (I) and / or formula (II), which may be synthesized manually or in a semi-automated or fully automated manner. In this case, it is possible, for example, to automate the procedure for producing such compounds, preparing or purifying the products or intermediates generally as described in, for example, by SH DeWitt in Annual Reports in Combinatonal Chernistry and Molecular Diversity: Automated Synthesis ", Volume I, Verlag Escom 1997, pp. 69 to 77. In addition, compounds of formula (I) and / or formula (II) may be prepared in part or in whole by solid phase supported methods. To this end, Individual intermediate steps or all intermediate steps of synthesis or a synthesis adapted to suit the procedure in question are linked to a synthetic resin.Solid phase supported synthesis methods are described extensively in the literature, for example, Barry A. Bunin in "The CominatorialIndex", Academic Press, 1998.

During this specification, the word "comprising", or variations such as "comprising" or "comprising", shall be understood to include the inclusion of a particular element, whole or step, or group of elements, whole or step, but the exclusion of any other element, totality or step, or group of elements, totality or step.

In some specific embodiments, it is noted that the compounds

used in this invention are not bestatin. The compound used in the invention is not 1,10-phenanthroline. In other embodiments, the compound used in the invention is not 2,2'-bipyridine.

The invention will hereinafter be described by means of the following

Non-limiting Figures and Examples.

Examples Example 1

Evaluation of the Remodeling Event Mechanism Associated with Lice Egg Hatching:

The hatching mechanism of the lice egg was evaluated under a

dissecting microscope. Female body lice were fed for an hour and a half to a rabbit before being transferred to a petri dish containing human hair. The petri dish was then placed in a 32 ° C incubator; 32% relative humidity. Within 5 hours of feeding, female lice began to lay their eggs. Each female lays up to 10 eggs in one session. The eggs developed over the next 7-9 days. Within the last 12 hours before hatching, the following changes were observed. The developing embryo's eyes could be clearly detected within the egg, with the developing embryo oriented so that its head is adjacent to the hatch flap or operculum. The embryo can be observed moving within the egg. The hatching occurs when the operculum opens and soon after the embryo begins to emerge. The head appears first, followed by the chest to which the legs are attached. Finally, the abdomen comes out and the nymph gets rid of the egg that remains stuck in her hair. There are no obvious structures associated with the newly emerged nymph's head visible under a light microscope that would facilitate hatching (ie no egg teeth are present). This observation suggests that while physical movement of the nymph within the egg probably contributes to egg hatching, other specific biochemical events are involved.

Example 2 Detection of Protease Activity in Lice Egg Extracts:

Within 12 hours of hatching, 50 body lice eggs (Pediculus humarius humanus) were removed from the hair and placed in a 1 mL eppendorf tube. 20 μΐ of distilled water was added to the un hatched eggs and the preparation incubated for 30 minutes at 32 ° C. The 20 µL was recovered, lyophilized and stored at -70 ° C. This sample was referred to as sample 1. A number of other samples were also collected as described. Sample 2 was collected by removing the un hatched four-strand louse eggs that were approximately 3 cm long, cutting the hair by 0.5 cm in length, and placing them in a microcentrifuge tube containing 20 cm. μΐ of distilled water and incubating at 32 ° C for 30 minutes. Sample 3 was collected as for sample 2, but the hair was placed in a tube containing 10 mL of 1% sodium hypochlorite for 1 minute, followed by five 1 minute washes in 25 mL of distilled water to remove hypochlorite. before being incubated in a microcentrifuge tube containing 20 μ of distilled water and incubated as for Sample 2. Sample 4 was collected from non-hatched eggs that were removed from the hair and washed with 1% sodium hypochlorite and incubated. in 20 μΐ of distilled water in the same manner as hair in Sample 3. Finally, Sample 5 was collected from eggs that were within 24 hours of hatching and washed with 1% sodium hypochlorite, then returned to incubator at 32 ° C until they hatched, the empty eggshells collected 0 to 2 hours after hatching, placed in a 1 mL microcentrifuge tube containing 20 μΐ of distilled water and incubated as for Samples 1- 4 For all 1-5 samples, the 20 μΐ Shy were collected, lyophilized and stored at -70 ° C. Refers to the washes recovered from these newly hatched eggshells as eggshell washes (ESWs). In order to look for proteases present in these different samples, the lyophilized samples were resuspended in 15 μΐ of non-reducing SDS sample buffer, centrifuged at 10,000g for 2 minutes and all 15μL loaded to 10% SDS-PAGE gels with gelatin substrate. The gels were fractionated at 4 ° C for 10 minutes at 10mA, followed by another 25 minutes at 15mA per gel. They were then incubated for 2 x 20 minutes in a 2.5% Triton-X 100 solution, followed by a three hour incubation in 0.1M Tris / HCl containing CaMb ImM pH 8.0. Activity was detected as clear areas in the gel, the result of protease activity degrading gelatin within the gel.

Protease activity on ESWs of louse eggs was also examined using two-dimensional gel electrophoresis. Large numbers of newly hatched eggshells need to be collected.

Following egg laying on pieces of cloth, the adult female lice were removed and the cloths with the eggs washed with 1% sodium hypochlorite as for the collection of ESWs. The eggs were then returned to the incubator and allowed to hatch. Typically 100 to 500 hatched eggshells were collected (0-2 hours after hatching), placed in a microcentrifuge tube containing 200 μΐ of distilled water, incubated and treated samples as described above. For analysis, ESWs were resuspended in rehydration buffer (8 M Urea, 2% 3 - [(3-cholamidopropyl) dimethylammonium] -1-propanesulfonate 2% pH-10 immobilized gradient buffer (Amersham Phamacia Biotech, Uppsala, Sweden) ) and used to rehydrate 7 cm Immobiline DryStrips pH 3-10 strips (Amershaw Bioscience) overnight. The strips were transferred to the Multiphor II apparatus (Pharmacia, Sweden), first electrophoresed at 200 V for 1 minute, increasing to 3,500 V over the next 90 minutes, followed by 65 minutes at 3,500 V, balanced (õ Urea, 30% glycerol, 50 mM Tris pH 8.8 and 2% SDS) and then fractionated on a 10% SDS-PAGE gel containing 0.1% gelatin to the second dimension. The gel was then fractionated and developed as previously described.

SDS-PAGE gelatin was used to analyze protease activity in ESWs of lice eggs both before and after hatching, and of human hair samples. An attempt was made to determine protein levels in each of the different ESW samples, however this proved unsuccessful due to the very low protein levels shown. Thus, for comparison purposes, samples were described in terms of the number of louse eggs from which washes were obtained.

Protease activity was detected in non-hatched egg washes within 12 hours of hatching (Sample 1) in the highest molecular weight region of the gel above 50 kDa (Figure IA, Column 1). A similar pattern of protease activity was detected in washes taken from human hair samples, followed by the removal of louse eggs (Sample 2) (Figure IA, Column 2). However, treatment of the hair with 1% sodium hypochlorite prior to wash collection (Sample 3) completely removed protease activity (Figure IA, Column 3). Hypochlorite treatment was also able to remove foreign proteases from un hatched louse eggs (Sample 4) (Figure IA, Column 4). Hypochlorite was used to treat un hatched eggs prior to ESW collection for all subsequent protease analyzes.

Several distinct proteases were observed in egg ESWs

treated with hypochlorite collected within 2 hours of hatching (Sample 5) (Figure 1B). Protease activity ranges were detected at about 25-30 kDa, 50 kDa and there were lighter ranges detected above 50 kDa.

SDS-PAGE in two-dimensional gelatin was used to further evaluate

precisely the number of protease species present in louse ESWs. Each of the three major regions of protease activity in one-dimensional gelatin SDS-PAGE (Figure 1B) is determined at a number of distinct proteases when analyzed by two-dimensional gelatin SDS-PAGE (Figure 3A). The proteases present in louse ESWs with activity in the 25-30 kDa molecular weight range determined in at least seven distinct proteases with isoelectric points in the neutral to alkaline pH range, while the protease activity range of about 50 kDa determined from at least eleven distinct protease regions with isoelectric points in the acid to neutral p1-1 region. The regular band pattern of proteases in the 50 kDa region suggests that they may be related in some way. At least five proteases with molecular weights above 75 kDa were also observed.

In conclusion, the hatching process in lice was studied by

light microscope. Egg hatching appears to be associated with physical activity of the developing nymph within the egg. However, the lack of any specialized structures to puncture or release the hatch flap or operculum indicates that the hatching process may also involve a biochemical component.

Although highly active proteases have been detected during egg hatching time in lice, the primary source of these proteases appears to be of maternal origin. Removal of this activity before hatching was achieved using sodium hypochlorite with the louse progressing to hatch successfully. Subsequent ESW analysis of newborn lice indicated the presence of a number of protease species that were further investigated as targets to inhibit egg hatching in lice.

Similar evaluations can be made for other pest egg remodeling events.

Example 3 Characterization of Proteases in Eggshell Washings:

In order to evaluate the potential of lice hatching proteases in eggshell washes as targets to inhibit egg hatching it was first necessary to characterize the nature of hatching proteases. Inhibitors of the 4 largest protease classes were used to classify proteases in the ESW.

Gelatine gels as substrate for 10% SDS-PAGE were loaded with lyophilized eggshell washes of 100 lice eggs which were resuspended in 50 μΐ ^ non-reducing sample buffer with 10 μΐ ^ liquefied samples per streak. The gels were fractionated at 4 ° C for 10 minutes at 10 mA per gel, followed by other minutes at 15 mA per gel. Gels were then cut into strips and each strip incubated for 2 χ 20 minutes in a 2.5% Triton-X 100 solution containing a specific inhibitor. The inhibitors used were PMSF serine protease inhibitor (5 mM), 1,10-phenanthroline metalloprotease inhibitors (10 mM) and 10 mM EDTA (tetraacetic ethylenediamine acid), Pepstatin aspartic protease inhibitor (5 µΜ) and inhibitor cysteine E-64 (10μΜ). The gel strips were then incubated in 0.1M Tris / HCl containing ImM CaCb pH 8 containing the different protease inhibitors for 3 hours at 37 ° C, before staining in Coomassie blue and discolouring as previously described.

Incubation with metal chelating agents EDTA and 1,10-phenanthroline to inhibit metalproteases resulted in a reduction in protease activity compared to untreated controls (Figure 2A and 2B, respectively). In contrast, there was no apparent reduction in protease activity when ESWs were incubated with the serine / cysteine protease inhibitor PMSF (Figure 2B), the cysteine protease inhibitor E-64 (Figure 2B) or aspartic protease inhibitor pepstatin (data not shown).

In order to further investigate the effect of 1,10-phenanthroline on the activity of

protease of eggshell washes, the proteases were separated by two-dimensional gel electrophoresis and the gel incubated in the presence of 10 mM 1,10-phenanthroline. The results of these studies confirmed the inhibitory effect of this metalloprotease inhibitor on louse egg protease activity. There was a general reduction in protease activity in the 25-30kDa region and a clear reduction in proteases around the 50kDa region and above 75kDa (Figure 3B).

A similar approach can be used to characterize proteins in eggshell washes of other thin membrane eggs of different pests.

Example 4

Development of an In Vitro Bioassay to Measure Lice Egg Hatching:

To evaluate the potential effects of protease inhibitors on lice egg closure, it was necessary to develop a reliable in vitro bioassay. Male and female body lice were fed on a rabbit as previously described. Female and male adult lice in a 3: 1 ratio were then transferred to a clean Petri dish containing approximately 3x3 cm2 nylon cloth and left for 12 hours at 32 ° C. During this time, the female louse laid its eggs and attached them to the tissue. All lice would then be removed and the eggs allowed to incubate for the next 5 days. On Day 6, the cloth containing the eggs would be placed for 1 minute in a 1% sodium hypochlorite solution and then washed thoroughly. The eggs would then progress to their final stages of development and hatch. In untreated control eggs, a reliable mean hatching percentage of between 85 and 95 percent was obtained using the in vitro egg hatching assay. It was subsequently discovered that for the egg hatching assay it was not necessary to pretreat the lice eggs with sodium hypochlorite.

A similar approach can be used to develop in vitro bioassays to measure other remodeling events that include, but are not limited to, thin membrane egg hatching. Example 5

Identification of Compounds That May Inhibit

Lice Hatching Protease Activity:

(a) Protease inhibitor test using lice egg hatching bioassay:

Having improved a bioassay to measure hatching eggs in eggs, the next phase of research was to use this bioassay as a means of testing the effects of different protease inhibitors on hatching eggs.

Lice eggs were clothed as described above. Cin-

Within days of placement, the cloth containing louse eggs was removed and immersed in a 1% sodium hypochlorite solution before being washed extensively in distilled water and dried with tissue paper. Lice eggs were counted under a dissecting microscope and the cloth cut into batches of between 10 and 30 eggs with 3-5 replicates used for treatment. The cloth containing louse eggs was then immersed in a protease inhibitor solution for a period of 10 minutes, placed on tissue paper for 1 minute to dry before being transferred to a clean Petri dish and incubated until hatching. . Eggs were observed at regular time intervals for evidence of eggs hatching over the next 1-2 days, by which time control eggs had hatched. Protease inhibitor solutions were typically prepared as stock solutions and added fresh at the appropriate concentration. A stock solution was specifically prepared as follows: 1,10-phenanthroline (200mM in methanol). In addition, equivalent solvent levels were added to control eggs containing no test inhibitor for any buffer isolated effects. The hatching inhibition percentage was calculated as the egg hatching reduction percentage compared to the untreated control. The untreated control was given a hatching percentage of 100%.

Addition of 1,10-phenanthroline, a metal chelating agent and a metalloprotease inhibitor significantly inhibited egg hatching in lice by 10 mM, whereas in ImM the level of inhibition was approximately 30% compared with that of lice. controls (refer to Figure 4).

These results provide data on the effect of a specific metal chelating agent and metalloprotease inhibitor on hatching of lice eggs. It was noted, however, that when 1,10-phenanthroline was added within 24 hours of hatching, variable egg hatching inhibition was observed (data not shown). This variability in hatching inhibition could be due to a number of factors that relate to the specific stage of louse development. Furthermore, these studies indicated that it is very difficult to predict the exact hatching time and therefore choosing a single time point at which to treat eggs can be problematic when assessing the effects of a specific inhibitor on egg hatching. The in vitro assay system was therefore modified to account for this variability in head development.

(b) Time course experiment using in vitro hatching test:

A series of time course experiments was conducted as a means of evaluating lice egg hatching inhibitors. Eggs were clothed as described above and then at 24 hour intervals an inhibitor was added to a new egg group for eggs up to 120 hours after laying. The eggs were then incubated at 28 ° C for another 8 days to allow egg hatching. This method of assessing inhibitors more closely reflects the field situation where lice eggs will be in various stages of development.

The results of these studies are shown in Table 1. Significant inhibition by 1,10-phenanthroline has been demonstrated at varying concentrations during the course of hatching. A degree of concentration dependence was also observed with the inhibitory effects of 1,10-phenanthroline. The results indicate that time course experiments provide more reliable means of evaluating the effects of specific inhibitors on hatching of lice eggs. Table 1

Percentage of egg hatching inhibition following treatment with different concentrations of 1,10-phenanthroline at 24 hour intervals

after laying eggs

Time after egg placement (h) Inhibitor 24h 48h 72h 96h 120h 144h mM 1,10- phenanthroline 100 100 100 100 100 100 mM 1,10- phenanthroline 100 100 100 100 93 96 2.5 mM 1,10- phenanthroline 100 100 85 85 89 60

The results of the above studies indicate that lice hatching enzymes are metal-class proteases judging from the ability of the metal chelating agent and metalloprotease inhibitor 1,10-phenanthroline to inhibit their activity. In addition, this compound was able to significantly inhibit egg hatching in head lice at all time points examined with some evidence of a dose-dependent effect particularly when eggs were treated at lower concentrations around hatching time. 1,10-phenanthroline exerts its effects by its ability to chelate metal ions, preferably zinc and thus inhibiting zinc-dependent proteases.

Example 6

Identification of Suitable Metal Chelating Agents for Inhibiting Lice or Plutella Eggs:

A number of metal chelating agents with the potential to inhibit remodeling events associated with lice thin-hatched egg hatching were analyzed by comparing their percentage ovicidal activity on lice at different concentrations, their calculated logotype pLDso, Pref pLDso, Activity class, clogP and spring refractivity were determined. Results are shown in Table 2. VO

(Ν Λ

i— <

i Xi d H

MR IO LO IO IO 00 LO clogP 2.05 2.05 2.56 2.56 Activity Class O = Inactive 1 = Low 2 = Medium 3 = High CS CS Pref pLDso CS Λ CS V CM a Logito calculated pLDso CO co "o" 2.85 ~ co ~ Concentration mM O ι — i ι — o CO o τ — i Ovicidal activity% 100.00% 0.00% 100.00% 100, 00% Lice (L) Plutella (P) J Dl, J Ph Name 1,10-phenanthroline 4,7-phenanthroline 5,5'-dimethyl bipyridine 5,5'-dimethyl bipyridine OO ιο VO LO 00 LO νΟ LO MD LO VO LO VO LO VO LO VO LO VO LO (Μ CN CN CN CN CN CN CO CO CM CNI CN CO τ — Ι CM Λ CO Λ ^ V CO Λ CN ^ tr CM τ — Ι CO CO IN CO CO ■ sl- "tf- CO CN CO ι — I ο" O ι — I. — ι τ — I ο "O ι — ι CD r — I 98.00% 57.00% 100.00% 100.00% 0.00 % 100.00% 100.00% CU Cu CU -J Cu PH J CU ê Cu '-M CC S .S' -M CC <υ ΰ • 3 os <υ β '-M 03 <υ α' -Μ ο3 S .S FH '-Μ (ΰ «υ à rB 03 <υ d • s 3 -α C C χί S = I ^! S -υ ^ § -B σ -α • rmmI ♦ κ Τ3 .tl Γο ιο ^ ίο IO ^ ι Ώ ^ · VO VO & οο σι

VO ■ Φ COo ooo 00 VO 00 VO LO LO νΟ IO VO IO τ — Ι IN IN LO LO CO CO r — I CM CN CN CN iH ι — I O CO CO CO CN CN CN V CN V CO Λ CN V CN CO IN ί- IN 00 IN CO O • • * —ι <ο, —ι O-1 O ι — — rl <D, - (O ι-I 65 0.00% 0.00% 0.00% 0.00% i 0.00% 100% I 0.00% Q Ph 2 J Oh J Oh J CU - § 9 CX JQ 4,4'-dimethyl ι bipyridine 2 2,2'-bipyridine 2-benzylpyridine 2-phenylpyridine 2,2 ', 6,2 "-terpyridine 2,2', 6,2" -terpyridine 2,2'-Bis (4,5-dimethylimidazole)

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> Example 7

Phenanthroline Washed Egg Post-Treatment Effect:

An experiment was performed to determine if egg washing would affect the inhibitory activity of 1,10-phenanthroline (Table 3). A control group (5% methanol) was also established. The percentage of hatching inhibition was calculated as the percentage reduction in egg hatching compared to the untreated control. The untreated control was given a hatching percentage of 100%. The results of this experiment indicate that 1,10-phenanthroline is still highly effective for inhibiting hatching of lice eggs by washing eggs in water. In later stage eggs that are approaching egg hatching (day 5) the effects appear to reflect a concentration dependence similar to that observed when lower inhibitor concentrations were used. It was also noted that a proportion of 1,10-phenanthroline-treated eggs had embryos that appeared to develop normally, but failed to hatch. Table 3

Percentage of egg hatching inhibition following treatment with 10 mM 1-10 phenanthroline at 24 hour intervals after egg laying in lice. Lice eggs were treated with inhibitor for 10 minutes and left without washing or treatment for 1 minute and then

Let it hatch.

Time after placement (h) 24h 48h 72h 96h 120h Treated / not 100 washed 100 Treated / washed 100 100 100 97 62

Example 8

Body Louse Egg Hash Inhibition with 1-10 Phenanthroline:

Tests were conducted to determine whether metal chelating agent and metalloprotease inhibitor 1,10-phenanthroline could inhibit hatching (Pediculis humanus capitus) egg hatching as opposed to body louse. Head lice eggs were obtained by placing groups of both adult male 1-2 head and adult female 6-8 lice in separate wells in a 24-well Petri dish containing cotton cloth. The petri dish was transferred to a humid incubator at 32 ° C, 70% RH for 12 hours to allow the female louse to lay its eggs. After 12 hours, all adult lice were removed from the Petri dish wells and a series of time course experiments were conducted. One egg group (24h old) was treated for 10 minutes with 200 µl of a 10 mM 1,10-phenanthroline solution. A control group (ie, no inhibitor treatment) of eggs was also included. Eggs were removed from the inhibitor, dried on tissue paper, placed at 32 ° C, 70% RH and allowed to hatch. A second group of eggs (48 hours old) were treated as previously described and also left to hatch. This process was repeated at 24 hour intervals on head lice eggs up to 120 hours after placement. This method of inhibitor evaluation more closely reflects the field situation in which lice eggs will be at various stages of development in the head and allows inhibitory effects to be observed at these different stages of the parasite.

The results of the above studies indicate that 1,10-phenanthroline may significantly inhibit egg hatching in head lice (Table 4).

Table 4

Percentage of egg hatching inhibition following treatment with 10 mlví 1,10-phenanthroline 24 hours after egg laying in lice relative to control.

Days after placement I 2 3 4 5 Treated 100 87 88 100 100

These results strongly suggest that body lice are an effective model for analyzing the effects of protease inhibitors on hatching eggs of head lice.

25 Example 9

Lice Egg Hatching Inhibition

with Metal Chelators:

Experiments were conducted using two metal chelating agents, 2,2'-dipyridine and 6,6'-dimethyl-2,2'-dipyridine, which have clogP values of 1.56 and 2.55 respectively and molar refractivities of 46 cm3. / mol and 56 cm3 / mol, respectively, to determine their effects on hatching of lice eggs. These compounds were tested in the standard louse assay to determine their ovicidal effects (refer to example 5 for method used for inhibitor testing). The results of this study are shown in Tables 5 and 6.

Table 5

Egg hatching results following treatment with 2,2'-dipyridyl at 24 hour intervals after egg placement. Results are given for: N (number of eggs per replicate), I-I (number of eggs successfully hatched) and Ph (number of eggs partially hatched).

Replicas 24h 48h 72h 96h 120h N H Ph N H Ph N H Ph N H Ph N H Ph 1 7 O O 6 O 7 O 13 0 0 10 0 0 2 8 O 14 14 10 0 0 3 11 O O - - - 14 O O 10 0 0 13 0 0 Table 6

Egg hatching results following treatment with 6,6'-Dimentyl-2,2'-dipyridyl at 24 hour intervals after egg placement. Results are given for: N (number of eggs per replicate), H (number of eggs successfully hatched) and Ph (number of eggs partially hatched).

Replicas 24h 48h 72h 96h 120h 0 0 3 11 0 0 6 0 0 10 0 0 18 0 0 - - -

The results of these studies indicate that both 6,6'-Dimethyl-2,2'-dipyridyl and 2,2'-dipyridyl showed very strong ovicidal activity, with which hatching of egg lice was completely inhibited at all time points. examined. Both 6,6'-Dimethyl-2,2'-dipyridyl and 2,2'-dipyridyl are metal chelating agents and metalloprotease inhibitors that are uninterleaved.

Example 10

Comparative Evaluation of Commercial Lice Products

with 1,10-Phenanthroline:

The ovicidal properties of the three largest commercial head lice products were evaluated in the standard lice egg hatching assay. The three commercial head lice products were as follows:

1. KP-24® Nelso Laboratories, active ingredients 1% maldison (malation);

2. RID® Bayer, active ingredients, 1% pyrethrins; and

3. NIX® Pfizer, active ingredients, 1% permethrin.

These three products have been tested according to the manufacturer's recommendations. Groups of eggs (24 hours old) were treated with the different products according to the manufacturers' recommendations for the appropriate period of time (5-10 minutes), followed by a 1-2 minute rinse in 32 ° C water. ° C. One positive control (10 mM 1,10-phenanthroline) and two negative controls (untreated and 20% methanol) were also incorporated. After exposure to the different products, the eggs were rinsed with warm water at 32 ° C before being dried on tissue paper and placed at 32 ° C, 70% RH and allowed to hatch. A second group of eggs (48 hours old) were treated as previously described and also allowed to hatch. This process was repeated at 24 hour intervals on head lice eggs up to 120 hours after placement. This method of analyzing inhibitors more closely reflects the field situation where lice eggs will be at various stages of development in the head and allows inhibitory effects to be observed at these different stages of the parasite. The results of these studies are shown in Table 7. Table 7

Egg hatching results following treatment with 3 commercial head lice products, 10 mM 1,10-phenanthroline and controls at 24 hour intervals after egg placement. Results are given for: N (number of eggs per replica), H (number of eggs per replica)

hatched successfully) and Ph (number of partially hatched eggs).

NIX-Pfizer Replicas 24h 48h 72h 96h 120h N II Ph N H Ph N H Ph N H Ph N II Ph 1 16 12 2 9 7 0 18 3 3 12 8 3 19 12 3 2 10 4 3 6 2 3 10 3 3 15 7 5 18 8 7 3 10 7 2 9 4 3 17 5 7 - - - 36 21 5 RID-Bayer Replicas 24h 48h 72h 96h 12 Oh N II Ph N II Ph N H Ph N H Ph N II Ph 1 8 0 3 12 3 4 7 0 0 8 0 0 14 0 1 2 8 2 5 7 0 1 5 1 2 8 0 0 - - - 3 5 π o 1 r \ X \ J r \ W rt U 1 X 3 11 0 0 - - - KP24KP24 Replicas 24h 48h 72h 96h 120h 120h N H Ph N H Ph N H Ph N H Ph N H Ph 1 7 7 0 10 10 0 10 1 3 10 0 0 10 0 0 2 6 6 0 10 9 0 0 0 0 7 0 0 8 0 0 3 9 8 0 12 0 1 1,10-phenanthroline (10 mM) Replicas 24h 48h 72h 96h 120h N H Ph N H Ph N H Ph N H Ph N H Ph 1 13 0 0 5 0 0 7 0 0 10 0 0 9 0 0 2 9 0 0 15 0 0 7 0 0 10 0 0 6 4 0 3 - - - 8 0 0 9 0 0 - - - 7 1 0 Control (20% Methanol) Replicas 24h 48h 72h 96h 120h N H Ph N H Ph N II Ph N H Ph N II Ph 1 - - - 14 14 0 10 1 0 0 10 10 0 13 13 0 2 - - - 5 4 0 8 8 0 10 9 0 7 7 0 3 - - - - - 0 9 7 0 4 4 0 10 10 0 Control (untreated) Replicas 24h 48h 72h 96h 120h N FI Ph N H Ph N H Ph N H Ph N II Ph 1 10 9 0 11 11 0 25 24 0 10 8 0 20 20 0 2 20 18 0 8 7 0 10 10 0 11 10 0 20 18 0 3 - - - 8 8 0 - - - 10 10 0 - - -

Test results from 3 commercial pediculicides indicate that they exhibit inconsistent levels of ovicidal activity across different stages of egg lice hatching. While 1,10-phenanthroline compound was highly effective in inhibiting hatching of lice eggs.

Example 11 Evaluation of Additional Commercial Lice Products:

The ovicidal properties of the two largest lice products

Commercial head lice were evaluated in the standard hatching egg hatching assay. The 2 commercial head lice products were as follows: 1. Pronto Plus® Del Laboratories Shampoo, active ingredients 0.33% Pyrethrins; and

2. Ready Plus® Mousse Del Laboratories Shampoo, active ingredients 0.33% Pyrethrins.

These products have been tested according to the manufacturer's recommendations. Groups of eggs (24 hours old) were treated with the different products according to the manufacturer's recommendations for the appropriate time period (5-10 minutes), followed by a 1-2 minute rinse in water at 32 ° C. ° C. Two negative controls (untreated and 20% ethanol) were also incorporated. After exposure to the different products, the eggs were dried on tissue paper and placed at 32 ° C, 70% RH and allowed to hatch. A second group of eggs (48 hours old) were treated as described above and also left to hatch. This process was repeated at 24 hour intervals on head louse eggs up to 120 hours after placement. This method of inhibitor analysis more closely reflects the field situation where lice eggs will be at various stages of development in the head and allows inhibitory effects to be observed at these different stages of the parasite. The results of these studies are presented in Table 8. Table 8

Egg hatching results following treatment with 2 commercial head lice products and controls at 24 hour intervals after egg placement. Results are given for: N (number of eggs per replicate), H (number of eggs successfully hatched) and Ph (number of eggs partially hatched).

Ready Plus Shampoo Replicas 24h 48h 72h 96h 120h N H Ph N H Ph N II Ph N II Ph N II Ph 1 14 10 2 11 9 0 30 27 0 35 30 0 40 38 2 2 20 15 3 21 18 0 19 16 0 42 36 0 38 29 5 3 - Ready Plus Mousse Shampoo Replicas 24h 48h 72h 96h 120h N II Ph N H Ph N H Ph N II Ph N H Ph 1 10 8 O 18 15 0 47 31 9 63 8 34 51 7 40 2 15 13 O 10 6 0 30 14 8 29 5 10 50 8 30 o O -1 1 1 1 O - - - 34 13 17 21 1 15 31 1 17 Control (ethanol) Replicas 24h 48h 72h 96h 120h N H Ph N H Ph N H Ph N H Ph N H Ph 1 12 10 O 18 16 0 40 36 1 21 20 0 49 47 0 2 11 9 O 21 18 0 41 37 0 28 26 0 39 36 0 3 11 11 O 13 11 0 75 70 0 29 27 0 36 34 0 Control (untreated) Replicas 24h 48h 72h 96h 120h N H Ph N H Ph N H Ph N H Ph N H Ph 1 10 9 O 27 26 0 61 60 0 50 49 1 48 46 0 2 3

Test results of 2 commercial pediculicides indicate that they exhibit very poor and inconsistent ovicidal activity through the different stages of egg lice hatching.

Example 12

Evaluation of Compounds in Egg Hatching of Plutella xylostella:

Hundreds of Plutella xylostella (Waite strain) eggs were collected and laid over a period of 24 hours. Within 3-5 hours of collection, eggs were treated with different inhibitors as described below.

Lots of Plutella eggs that were laid on a thin cloth or paraffin were dipped in a specific inhibitor solution for between 2 and 10 seconds, the excess solution drained away by drying with tissue paper. The set of eggs was then placed in a humid box at 25 ° C until hatching. Control eggs were exposed to absolute methanol as described above. On day 6 after laying, eggs were evaluated for the different treatments and egg hatching percentage was determined relative to control, as shown in Table 9. Table 9

Ovicidal effects of inhibitors on hatching eggs of Plutella xylosíella in

relation to control.

Inhibitor Number hatched Number not hatched% Inhibition 6,6'-dimethyl-2,2'-dipyridyl (10 mM) 0 79 100 6,6'-dimethyl-2,2'-dipyridyl (1 mM) 0 26 100 6,6'-dimethyl-2,2'-dipyridyl (O1ImM) 23 29 0 6,6'-dimethyl-2,2'-dipyridyl (0.01mM) 13 7 0 6,6'-dimethyl-2,2 '-dipyridyl (0.001mM) 11 6 0 1,10-phenanthroline (10 mM) 0 45 100 1,10-phenanthroline (ImM) 15 16 0 Control (100% MeOH) 63 92 -

Table 9 indicates that 6,6'-dimentyl-2,2'-dipyridyl metal chelator was able to inhibit egg hatching in Plutella xylostella in a dose-dependent manner, with strong ovicidal effects evident in both 10 and Imm. In addition, the metalloproteinase / metal chelator inhibitor 1,10-phenanthroline was also able to significantly inhibit egg hatching of this insect by 10 mM. Example 13 Evaluation of Compounds in Plutella xylostella Egg Hatching:

Hundreds of Plutella xylostella (Waite strain) eggs were collected and laid over a period of 24 hours. Within 3-5 hours of collection, eggs were treated with different inhibitors as described below.

Lots of Plutella eggs that were laid on a thin cloth or paraffin were dipped in a specific inhibitor solution for between 2 and 10 seconds, the excess solution drained away by drying with tissue paper. The egg set was then placed in a 25 degree wet box until the egg hatched. Control eggs were exposed to absolute methanol as described above or untreated. On day 6 after laying, eggs were evaluated for different treatments and egg hatching percentage was determined relative to controls, as shown in Tables 10 and 11. Table 10

Ovicidal effects of inhibitors on hatching of Plutella xylosíella eggs in relation to controls (eggs laid on cloth).

Inhibitor Number hatched Number not hatched% Inhibition 6,6'-dimethyl-2,2'-dipyridyl (10 mM) 0 53 100 6,6'-dimethyl-2,2'-dipyridyl (ImM) 0 23 100 6 6'-dimethyl-2,2'-dipyridyl (OlImM) 49 0 0 6,6'-dimethyl-2,2'-dipyridyl (0.01mM) 23 4 12 5,5'-dimethyl-2,2 ' -dipyridyl (10 mM) 0 21 100 5,5'-dimethyl -2,2'-dipyridyl (ImM) 5 22 78 4,4'-dimethyl-2,2'-dipyridyl (10 mM) 0 36 100 4, 4'-dimethyl-2,2'-dipyridyl (ImM) 0 30 100 Control (untreated) 32 1 - Control (100% MeOH) 34 1 - Table 11

Ovicidal effects of inhibitors on hatching of Plutella xylostella eggs in relation to controls (eggs laid in parafilm).

Inhibitor Number hatched Number not hatched% Inhibition 6,6'-dimethyl-2,2'-dipyridyl (10 mM) 0 106 100 6,6'-dimethyl-2,2'-dipyridyl (ImM) 0 63 100 6 2,6'-dimethyl-2,2'-dipyridyl (OlImM) 65 11 7 6,6'-dimethyl-2,2'-dipyridyl (O1OmM) 92 1 0 5,5'-dimethyl-2,2'-dipyridyl (10 mM) 0 138 100 5,5'-dimethyl-2,2'-dipyridyl (1 mM) 18 133 88 4,4'-dimethyl-2,2'-dipyridyl (10 mM) 0 139 100 4.4 '-dimethyl-2,2'-dipyridyl (1 mM) 10 107 91 Control (untreated) 108 3 - Control (100% MeOH) 58 7 -

Tables 10 and 11 show the effects of exposure of Plutella xylostella eggs to selected dipyridyl egg hatching compounds relative to controls. The results show a dose-dependent effect for 6,6'-dimethyl-2,2'-dipyridyl with both 10 and ImM being effective in inhibiting hatching of Plutella eggs. At 0.1 and 0.01mM, there were no observable effects on egg hatching. These results confirm the results shown in Example 12 for this compound. In addition, both 5,5'-dimethyl-2,2'-dipyridyl and 4,4'-dimethyl-2,2'-dipyridyl were able to significantly inhibit egg hatching in both 10 and ImM.

There were no significant differences observed between laid eggs.

cloth or parafilm.

Example 14

Helicoverpa control spp. With Industrial Standard:

Lannate® (Crop Care Australasia Pty Ltd) containing metomil as an active compound was chosen as a comparative control.

For field control of Cotton Helicoverpa an application rate of 200mL / 100L is recommended. This equates to approximately 2.5 mM active compound. A range of concentrations was composed in water and the eggs placed in the solutions for approximately 2-10 seconds. The eggs that had been clothed were then dried with absorbent paper and placed in an incubator at 26 ° C for 3-4 days until hatching. Hatching rates were then evaluated compared to the water-only control (Figure 5). The results indicate a strong dose titration effect of Lanna ™ against Helicoverpa eggs with very good efficacy evident at 1.25 mM.

Example 15

Helicoverpa control spp. with 2-Acetyl-1-Tetralone:

The ovicidal activity of 2-acetyl tetralone against H. armige eggs was evaluated as described in Example 14. The compound was dissolved in 100% diethoxy glycol and diluted to a final concentration of 1% in water. The results are given in Figure 6 and showed that 2-acetyl-1-tetralone showed strong ovicidal efficacy in 2mM with decreased efficacy in ImM.

Example 16 Evaluation of Compounds in the Egg Hatching of Helicoverpa armigera:

Hundreds of Helicoverpa armigera eggs (Tatura χ Toowoomba strains) were collected and laid on fine mesh cloth over a 24-hour period. Within 3 to 5 hours of collection, all eggs were treated with different inhibitors as described below.

Lots of Helicoverpa eggs that were exposed to a specific inhibitor solution for between 2 and 10 seconds, the excess solution was drained by drying with tissue paper. The egg set was then placed in a 25 degree wet box until the egg hatched. Control eggs were exposed to absolute methanol as described above. On day 6 after laying, eggs were evaluated for the different treatments and egg hatching percentage was determined relative to control, as shown in Table 12. Table 12

Ovicidal Effects of Inhibitors on Egg Hatching of Helicoverpa Eggs

armigera in relation to the control.

Inhibitor Number hatched Number not hatched% Inhibition 6,6'-dimethyl-2,2'-dipyridyl (10 mM) 7 98 94 6,6'-dimethyl-2,2'-dipyridyl (1 mM) 4 140 97 6,6'-dimethyl-2,2'-dipyridyl (0.1 mM) on 0,6,6'-dimethyl-2,2'-dipyridyl (O 1 O 1 m) on 0,6,6'-dimethyl-2, 2'-dipyridyl (0.001mM) in the Na 1,10-phenanthroline (10 mM) 31 16 44 1,10-phenanthroline (ImM) in the aria. Control (100% MeOH) at na 0

na refers to all eggs hatching and being devoured by newborn caterpillars.

The results in Table 12 indicate that 6,6'-dimethyl-2,2'-dipyridyl was able to significantly inhibit egg hatching from Helcoverpa armigera eggs at 10 and ImM. No inhibitions were recorded at concentrations below this level. The 1,10-phenanthroline compound was also able to inhibit egg hatching by 10 mM only.

Example 17

Evaluation of Compounds in the Egg Hovering of Helicoverpa armigera:

Hundreds of Helicoverpa armigera eggs (Tatura χ Toowoomba strains) were collected and laid on fine mesh cloth over a 24-hour period. Within 3-5 hours of collection, all eggs were treated with different inhibitors as described below.

Lots of I-Ielicoverpa eggs were then exposed to a specific inhibitor solution for between 2 and 10 seconds, the excess solution drained by drying with tissue paper. The set of eggs was then placed in a humid box at 25 ° C until hatching. Control eggs were exposed to absolute methanol as described above. On day 6 after laying, eggs were evaluated for the different treatments and egg hatching percentage was determined relative to control, as shown in Table 13. Table 13

Ovicidal Effects of Inhibitors on Egg Hatching from Helicoverpa armigera

in relation to control.

Inhibitor Number hatched Number not hatched% Inhibition 6,6'-dimethyl-2,2'-dipyridyl (10 mM) 3 48 94 6,6'-dimethyl-2,2'-dipyridyl (ImM) 2 61 97 6 2,6'-dimethyl-2,2'-dipyridyl (0.1mM) 70 0 0 5,5'-dimethyl-2,2'-dipyridyl (10mM) 0 42 100 4,4'-dimethyl-2,2 '-dipyridyl (10 mM) 8 43 84 4,4'-dimethyl-2,2'-dipyridyl (ImM) 23 29 66 Control (100% MeOH) 37 2 -

The data presented in Table 13 support the previous results.

provided in Example 4, demonstrating that 6,6'-dimethyl-2,2'-dipyridyl is capable of significantly inhibiting the egg hatching of Helicoverpa armigera eggs in both 10 and ImM. In OjImM no inhibition of egg hatching with this compound was observed. In addition, data are presented indicating significant inhibition of egg hatching in mM for both 5,5'-dimethyl-2,2'-dipyridyl and 4,4'-dimethyl-2,2'-dipyridyl. In addition, significant inhibition of egg hatching was observed in ImM of 4,4'-dimethyl-2,2'-dipyridyl.

Example 18

15

Evaluation of Effects of 2- (2-Pyridinyl) Quinone on Plutella xylostella Egg Hatching:

Hundreds of Plutella xylostella (Waite strain) eggs were collected and laid over a period of 24 hours. Within 24-48 hours of collection, eggs were treated with different inhibitors as described below.

Lots of Plutella eggs that were laid on a thin cloth were dipped in a specific inhibitor solution for approximately 2 seconds, the excess solution drained by drying with tissue paper. The egg set was then placed in a 25 degree wet box until the egg hatched. Control eggs were exposed to absolute ethanol as described above. On day 6 after laying, eggs were evaluated for different treatments and egg hatching percentage was determined relative to control.

Table 14

Ovicidal effects of inhibitors on hatching of Plutella xylostella eggs in

relation to control.

Inhibitor Number hatched Niliiictu not hatched% Inhibition 2- (2-pyridinyl) quinoline (10 mM) 2 56 96 Control (100% ETOH) 55 14 -

Table 14 indicates that the 2- (2-pyridinyl) quinoline metal chelating compound was able to inhibit egg hatching in 10 mM Plutella xylostella.

Example 19

Evaluation of Effects of Added Metal Ions on 6,6'-Dimethyl-2-29-Dipyridyl Egg Hatching Inhibition: Hundreds of Plutella xylostella (Waite strain) eggs were collected and laid over a period of 6 months. 24 hours. Within 24 hours of collection, the following experimental model was chosen. Lots of eggs were exposed to 10 mM 6,6'-dimethyl-2,2'-dipyridyl for 2 seconds, while additional batches of eggs were exposed to solvent only (Methanol) for 2 seconds. All batches of eggs were allowed to air dry for 20 minutes at room temperature. The eggs were then exposed for 2 seconds to FeS04 at 10, 5 or ImM, air dried and placed in the 24 ° C incubator and allowed to hatch over the next 6 days. In addition, a 10 mM 6,6'-dimethyl-2,2'-dipyridyl positive control was created, in which eggs were exposed to this compound for 2 seconds, air dried and placed in the incubator. Table 15

Reversal of the 10 mM ovicidal effects of 6,6'-dimethyl-2,2'-dipyridyl on hatching of Plutella xylostella eggs compared to FeSCU controls.

Inhibitor Number hatched Number not hatched% Inhibition 6,6'-dimethyl-2,2'-dipyridyl (+ ve) control 0 44 100 6,6'-dimethyl-2,2'-dipyridyl followed by 2 second exposure MEOH 3 28 90 6,6'-dimethyl-2,2'-dipyridyl followed by 2 second exposure to 10 mM FeSÜ4 12 19 38 6,6'-dimethyl-2,2'-dipyridyl followed by 2 second exposure at 5 mM FeS04 25 0 0 6,6'-dimethyl-2,2'-dipyridyl followed by 2 second exposure to ImM FeS04 η Λ 1 3

The results presented in Table 15 indicate that the addition of

of the divalent Fe-like metal ions in FeSÜ4 was able to reverse the effects of the 6,6'-dimethyl-2,2'-dipyridyl metal chelating agent.

The results indicate that the reversal of the inhibitory effects of 6,6'-dimethyl-2,2'-dipyridyl is due to Fe replacing the action of this inhibitor as opposed to a simple dilution of the inhibitor with FeS04. This effect is indicated by the finding that exposure of eggs to MEOH alone after inhibitor exposure still resulted in a significant degree of egg hatching inhibition.

Example 20

Evaluation of Effects of Added Metal Ions on Egg Hatching Inhibition by 5,5'-Dimethyl-2,2'-Dipyridyl:

Hundreds of Plutella xylostella (Waite strain) eggs were collected and laid over a period of 24 hours. Within 24 hours of collection, the following experimental model was chosen. Lots of eggs were exposed to 10 mM 5,5'-dimethyl-2,2'-dipyridyl for 2 seconds, while additional batches of eggs were exposed to solvent only (Methanol) for 2 seconds. All batches of eggs were allowed to air dry for 20 minutes at room temperature. The eggs were then exposed for 2 seconds to FeS04 at 10, 5 or ImM, air dried and placed in the 24 ° C incubator and allowed to hatch over the next 6 days. In addition, a 10 mM 5,5'-dimethyl-2,2'-dipyridyl positive control was created, in which eggs were exposed to this compound for 2 seconds, air dried and placed in the incubator. Table 16

Reversal of 10 mM 5,5'-dimethyl-2,2'-dipyridyl ovicidal effects on hatching of Plutella xylostella eggs compared to FeSC controls> 4

only.

Inhibitor Number hatched Number not hatched% Inhibition 5,5'-dimethyl-2,2'-dipyridyl (+ ve) control 0 38 100 5,5'-dimethyl-2,2'-dipyridyl followed by 2 second exposure MEOH 16 19 55 5,5'-dimethyl-2,2'-dipyridyl followed by 2 second exposure at 10 mM FeSÜ4 23 2 8 5,5'-dimethyl-2,2'-dipyridyl followed by 2 second exposure at 5 mM FeS04 25 0 0 cr cr? Hypophilic O '' -J-M- o, -aimetii -ui ^ muiict followed by 2 second exposure to ImM FeS04 39 0 0

The results presented in Table 16 indicate that the addition of divalent Fe-ions metal ions in FeSÜ4 was able to reverse the effects of the 5,5'-dimethyl-2,2'-dipyridyl metal chelating agent. The results indicate that the reversal of the inhibitory effects of 5,5'-dimethyl-2,2'-dipyridyl is due to Fe removing the action of this inhibitor as opposed to a simple dilution of the inhibitor by FeS04. This effect is indicated by the finding that exposure of eggs to MEOFI alone after inhibitor exposure still resulted in a significant degree of egg hatching inhibition.

Example 21

Ovicidal Effectiveness of Metal Chelating Compounds

Against Plutella:

The ovicidal efficacy of 2-acetyl-1-tetralone has also been tested

brings eggs from Plutella. The compound was dissolved in diethoxy glycol and then diluted to a final concentration of 1% diethoxy glycol containing ImM 2-acetyl-1-tetralone and tested in the same manner as in Example 13. The results of this assay are given in Figure 7 and indicate strong efficacy. ovicidal resistance of this compound against Plutella at 2 and ImM without inhibition observed at 0.5 mM.

Example 22

Evaluation of Compounds in Egg Hatching

from Plutella xylostella:

Ovicidal efficacy He 2,2 ', 6,2 "-terpyridine and 5,5'-diethyl-2,2'-uipyridine

It has also been tested against Plutella eggs. The compounds were dissolved in diethoxy glycol and then diluted to a final concentration of 1% diethoxy glycol containing 1 mM 2,2 ', 6,2'-terpyridine or ImM and 0,1 mM 5,5'-diethyl-2,2' Dipyridyl and tested in the same manner as in Example 13. The results of this assay are given in Table 17 and indicate complete inhibition in ImM for both compounds.Inhibition of egg hatching was observed in O 5 µmM 5,5-diethyl. -2,2'-dipyridyl Table 17

Ovicidal effects of compounds on hatching of Plutella xylostella eggs in relation to controls (laid eggs).

Inhibitor Number hatched Number not hatched% Inhibition 5,5'-diethyl-2,2'-dipyridyl (1 mM) 0 34 100 5,5'-diethyl-2,2'-dipyridyl (0.1 mM) 22 34 31 2.2 ', 6.2 "-terpyridine (ImM) 0 46 100 Control (1% diethoxyglycol) 37 39 -

Example 23

Effects of 6,6'-Dimethyl-2,2'-Dipyridyl and 5,5'-Dimethyl-2,2'-Dipyridyl

at the Egg Hatching in Bovicola ovis:

B. ovis eggs were collected from the wool of sheep that were infested with this parasite. Eggs were collected using tweezers and with the aid of a dissecting microscope and placed in 24-well tissue culture plates in duplicate groups of 10 eggs per replicate. Eggs were then exposed to either methanol alone (control) or test compounds for either 10 minutes or 1 minute before being removed from the wells and placed in glass bottles containing nourishment at the base of the tube. The tubes were placed in plastic containers containing a salt solution (to maintain constant 68% humidity) and the containers kept at a temperature of 32 ° C. Eggs were monitored for hatching for the next 12 days and% hatching inhibition determined compared to controls. Table 18

Effects of 6,6'-dimethyl-2,2'-dipyridyl and 5,5'-dimethyl-2,2'-dipyridyl on egg hatching in Bovicola ovis.

Number of Hatches Inhibitor Number in different diffusion%% Inhibition of replicates given mM 5,5'-dimethyl-2,2'- Rep 1. 0 10 100 dipyridyl (10 minutes exposure) Rep 2. 0 10 mM 5 , 5'-dimethyl-2,2 Rep 1. 0 10 100 dipyridyl (1 minute exposure) Rep 2. 0 10 mM 6,6'-dimethyl-2,2'-Rep 1. 0 10 100 dipyridyl ( 10 min exposure) Rep 2. 0 10 mM 6,6'-dimethyl-2,2'- Rep 1. 0 10 100 dipyridyl (1 min exposure) Rep 2. 0 10 Control (Ethanol) (10 min Rep 1. 5 5 - Exposure Rep 2. 5 5 Control (Ethanol) (1 minute Rep 1. 4 6 - Exposure) Rep 2. 5 5 Rep 1. 3 7 - Control (Untreated) Rep 2. 6 3

The results presented in Table 18 indicate that following a 10 or 1 minute exposure of B. bovis louse eggs to a 10 mM solution of 5,5'-dimethyl-2,2'-dipyridyl or 6,6'-dimethyl 2,2'-dipyridine Egg hatching in this ectoparasite could be completely inhibited in this assay.

Example 24

Effects of Metal Chelating Agents on Egg Hatching in Haemonchus contortus:

The gastrointestinal parasite Haemonchus contortus is a major sheep pathogen worldwide. The parasite survives through the ability of adult worms to attach to the sheep's abomasal mucosa and draw blood. An adult female can draw approximately 0.1ml of blood per day. Adults can live for many months with females producing hundreds of eggs per day and infected animals dropping more than 1,000 eggs per gram of feces per day on pasture. Eggs hatch after 1-2 days depending on weather conditions and, following two seedlings, infectious L3 larvae appear on pasture and are consumed by the host. Once in the host, L3 larvae spill from the sheath into the rumen. they migrate to the abomasum and start feeding by burrowing in the mucosa, where they progress for yet 2 seedlings. The infected sheep have their conditions weakened and in more severe cases suffer dehydration and anemia due to blood loss. If the parasites are not removed, the animals will die. Control is centered on the strategic use of anthelmintics along with pasture management. Growing problems with parasite resistance pose significant problems for producers, as most anthelmintics on the market are no longer effective against this parasite. In fact, even ivermectin, which had shown significant potency to control this parasite, began to fail to develop resistance. In an attempt to improve H. contortus control, the effects of 5,5'-dimethyl-2,2'-dipyridyl were examined in eggs II. contortus Eggs were recovered from the infected sheep feces using a standard sucrose flotation method. The eggs were set to a density of approximately 2,500 eggs per mL and were then exposed to varying concentrations of the compound and incubated for 48 hours at 25 ° C. Eggs were then examined to determine the ovicidal efficacy of the compound compared to untreated control and a solvent only control. Figure 8 shows the ovicidal efficacy of 5,5'-dimethyl-2,2'-dipyridyl in eggs of II. contortus

Figure 8 indicates that 5,5'-dimethyl-2,2'-dipyridyl was potentially ovicidal at 180 and 18 µg / mL (equivalent to 1 and 0.1 mM of the active, respectively). A comparison of the ovicidal efficacy between 5,5'-dimethyl-2,2'-dipyridyl and the commercial product ivermectin indicated that 5,5'-dimethyl-2,2'-dipyridyl was in the order of 10x most effective in hatching inhibition. contortus eggs compared to ivermectin (Figure 8 and Figure 9).

In addition, the ovicidal efficacy of the 2-acetyl-1-tetralone compound was examined against H. contortus (Figure 10). The same protocol was used as described for 5,5'-dimefil-2,2'-dipyridyl. The data indicate that at 220ug / ml (ImM) and 10ug / ml (0.5mM) 2-acetyl-1-tetralone was highly effective inhibiting egg hatching. Partial inhibition of egg hatching was observed at 22ug / mL (0.1mM).

Example 25 Effects of 5,5'-Dimethyl-2,2'-Dipyridyl on Egg Hatching and Viability

in Dermatophagoides spp mites:

A filter paper (90 mm in diameter) was picked up and placed in a petri dish of the same dimensions. The filter paper was then thoroughly wetted with the test compound using a small air pump sprayer. The wet filter paper was then allowed to dry in the free air flow. 200mg of mite medium, containing approximately 500 mites / g, was placed on filter paper and the number of mites counted under a microscope. The arena was then left for 2 weeks in an incubator at 25 ° C and 75% RH. After 2 weeks, the number of mites in the arena was counted a second time. This experiment was repeated 3 times with the test compound and another 3 times using water as a control. Results are presented in table 19.

Results

Replica 1 Hour Count 2 Week Count 1 Test Compound 76 74 2 Test Compound 61 50 3 Test Compound 104 90 4 Control 69 110 Control 72 125 6 Control 91 112

Table 19

Mite count after 1 hour and two weeks on test papers treated with control compound

The mite populations in treated filter papers showed

a slight decline over the 2 week period. This may be due to egg hatching prevention and / or oviposition effects of female mites. In contrast, control treatments showed small decreases in mite populations, suggesting a lower mortality of adult mites and no adverse effect on egg viability.

Example 26 Effects of 5,5'-Dimethyl-2,2'-Dipyridyl on Egg Hatching on Cat Fleas Ctenocephalides felis.

Cat flea eggs were exposed for 10 minutes to 10 mM 5,5'-dimethyl-2,2'-dipyridyl and then removed from solution and placed in an incubator and allowed to hatch. A group of mite eggs was exposed to the vehicle only and was used as a control. A third group remained untreated. Subsequently, the eggs were examined and the percentage of eggs that hatched successfully compared to the given controls.

Example 27 Effects of 5,5'-Dimethyl-2,2'-Dipyridyl on Egg Hatching in Cimex lectularius Bedbugs:

Bedbug eggs were exposed for 10 minutes to 10 mM 5,5'-dimethyl-2,2'-dipyridyl and then removed from solution and placed in an incubator and allowed to hatch. A group of mite eggs was exposed only to the vehicle and was used as a control. A third group remained untreated. Subsequently, the eggs were examined and the percentage of eggs that hatched successfully compared with the given controls.

Example 28 Effects of 5,5'-Dimethyl-2,2'-Dipyridyl on Haemonchus contortus Survival: Third stage II larvae were exposed. contortus at varying concentrations of 5,5'-dimethyl-2,2'-dipyridyl and the molting effects of L3 and L4 examined. Larvae had their sheaths shed (their L2 sheaths were chemically removed) or not shed (their L2 sheaths were intact). Larvae were exposed to varying concentrations of 5,5'-dimethyl-2,2'-dipyridyl added to their DMEM culture medium and the effects on larval survival monitored over time. Following a 30 minute incubation at 37 ° C, more than 90% of the spilled larvae exposed to 0.5 mM of the compound appeared dead. In contrast, no adverse effects were observed on spilled larvae compared to control larvae within 3 days of exposure to the compound. On day six after exposure, more than 90% of the larvae appeared to have died in the treatment groups while the control larvae appeared healthy. This larvicidal effect on spilled larvae appeared to be dose dependent as following exposure of ΙΟμΜ of the compound the larvae appeared normal within 6 days of exposure.

Example 29

Effect of 5,5'-Dimethyl-2,2'-Dipyridyl Formulated on Egg Lice Hatching:

5,5'-dimethyl-2,2'-dipyridyl was formulated and evaluated in the standard body louse egg assay.

Body louse eggs (15-30 per replica) of varying ages were exposed for 10 minutes to the test solutions or to a placebo or left untreated, followed by washing with water for 1 minute and dried with absorbent paper. Eggs were then incubated at 30 ° C for the next 10 days after treatment and the percentage of eggs that hatched successfully was determined (Table 20). The results show a strong dose dependence of ovicidal activity when 5,5'-dimethyl-2,2'-dipyridyl compound is formulated and applied to body louse eggs.

Table 20

A summary of the data is presented below. Data are expressed as the% of eggs that hatched following treatment.

Treatment Egg Age 24h 48h 96h 120h Placebo Formulation 88 93 92 96 Control (Untreated) 89 84 93 95 5,5'-dimethyl-2,2'-dipyridyl (30mM) 0 0 0 0 5,5'-dimethyl -2,2'-dipyridyl (10 mM) 4 7 10 12 5,5'-dimethyl-2,2'-dipyridyl (5 mM) 21 58 64 65 5,5'-dimethyl-2,2'-dipyridyl ( 1 mM) 91 80 82 86

Example 30

Effect of Formulated S-Dimethyl-Dipyridyl

in Egg Lice Hatching:

Pregnant female lice were allowed to lay eggs. Eggs were counted, inspected under a light microscope and all eggs that appeared undamaged were allocated to one of two treatment groups. One group was exposed to 5'-dimethyl-2,2'-dipyridyl in one formulation, while the second group was exposed to the formulation only. The protocol was as follows:

Treatments: 5'-dimethyl-2,2'-dipyridyl (20mM) n = 10 Vehicle control only n = 10 Exposure time: 10 minutes Washing time: 1 minute at ~ 37 ° C

Incubation: All eggs were placed at 31 ° C in a humid incubator and monitored for signs of specifically eye development and subsequent hatching, see Table 21.

Results:

Table 21

Day 1 (03/09)

Ha44 Treatment Placebo Developmental stage 1 eye development 9 without des. eye development 0 eye development Day 4 (13/03) Ha44 Treatment Placebo Developmental stage 1 hatched 1 eye development 8 no des. Eye 4 Eye 6 development without des. of the eye Day 5 (3/14)

Ha44 Treatment Placebo Developmental stage 1 eye development 8 without des. of eye 8 development of eye 2 without des. Eye Treatment Day 6 (3/15) Ha44 Treatment Placebo Developmental Stage 1 Eye Development 8 without development. of eye 8 development of eye 1 without des. Eye Day 7 (16/03) Ha44 Treatment Placebo Developmental Stage 1 Eye Development 8 without development. 5 hatched 4 eye development 1 without des. Eye Day 12 (21/03) Ha44 Treatment Placebo Developmental Stage 1 Eye Development 8 without development. hatched 6 eye 3 eye development 1 without des. of the eye

Results indicate that a 20mM formulation of 5'-dimethyl-2,2'-dipyridyl can significantly suppress egg hatching in head lice. Two of the eggs developed, but only one of the two eggs hatched. The other 80% failed to hatch. In the Placebo-treated group, 60% of eggs hatched, 30% developed into nymphs, but did not hatch, and 1 egg did not develop. These data indicate that a formulation containing 5'-dimethyl-2,2'-dipyridyl may significantly inhibit hatching in head lice eggs.

Example 31

Ovicidal Effect of 5,5'-Dimethyl-2,2'-Dipyridyl and 5,5'-Diethyl-2,2'-Dipyridyl on Egg Hutting in Plutella Greenhouse:

A number of treatments and replicas have been established on the glass house using young cabbage plants that were in the 4-5 foliage stage. Plutella xylostella eggs were laid by pregnant females on the leaves, such that each plant contained 10 eggs. Plants were sprayed with a commercial ovicide or using the compounds 5,5'-dimethyl-2,2'-dipyridyl and 5,5'-diethyl-2,2'-dipyridyl in a formulation at the rate of 200 L / Ha using a track sprayer. Also included were water controls only or placebo. Ovicidal efficacy was monitored for a number of days and the number of hatching eggs and emerging caterpillar larvae was recorded (Table 22). (Μ (Ν

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It will be noted by persons skilled in the art that numerous

Variations and / or modifications may be made to the invention, as shown in the specific embodiments, without departing from the spirit or scope of the invention as broadly described. These modalities should therefore be considered in all respects as illustrative and not restrictive.

All publications discussed above are incorporated herein in their entirety.

Any discussion of documents, acts, materials, devices, articles or the like that has been included in this Descriptive Report is solely for the purpose of providing a context for the present invention. It should not be taken as recognition that any or all of these subjects are part of the prior art base or were common knowledge in the field relevant to the present invention as existed in any country prior to the priority date of each Claim of this Order.

Claims (42)

"Methods of Treatment of Plague Infestation, Invertebrate Plague Extermination, Remodeling Event Inhibition and Egg Incubation in Non-Ectoparasitic Invertebrates in Invertebrate Population, Control or Extermination of Invertebrate Plague Population Chelating Agent As Candidate Invertebrate Remodeling Event Inhibitor From Metal Chelating Agents Collection "Claims A plague infestation treatment method, characterized in that it comprises decreasing the exsheathment of an invertebrate by externally contacting a plague with a compound of formula (I): <formula> formula see original document page 146 < wherein X is selected from a covalent bond, -C (RS) 2-, -Z- or -C (RS) 2 -ZC (RS) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 , CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 52 -, -C (R 5) 2, -C ( R5) 2-, -CR5 = CR5-, C (O), C (S) or NH; R2, R2 ', R3, R3', R4 and R4 'are independently selected from hydrogen, C1-6 alkyl , a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered carbocyclic or heterocyclic ring; R3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a 5- or 6-membered carbocyclic or heterocyclic ring; independently selected from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2-C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or decrease the exsheathment rate of said invertebrate. 2. A plague infestation treatment method, characterized in that it comprises the decreasing excistion of an invertebrate by external contact of a plague with a compound of the formula: where X is selected from a covalent bond, -C (R5) 2-, -Z- or -C (RS) 2 -ZC (RS) 2-; R 1 and R 1 "are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 , CO 2 H, CO 2 C 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C (R5) 2-, -CR5 = CR5-, C (O), C (S) or NH; R2, R2 ', R3, R3', R4 and R4 'are independently selected from hydrogen, C1-6. 6alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered carbocyclic or heterocyclic ring, or R 2 and R 3 or R3 and R4 and / or R2 'and R3' or R3 'and R4' taken together with the carbon atoms to which they are attached form a (I) 5 or 6 membered carbocyclic or heterocyclic ring; R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-4 alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and acceptable thereof in an amount effective to inhibit or otherwise decrease the rate of excitation of said invertebrate. 3. A plague infestation treatment method, characterized in that it comprises decreasing the apolysis of an invertebrate by externally contacting a plague with a compound of the formula: Z is selected from a covalent bond, -NH-; > formula see original document page 149 </formula> a pharmaceutically, veterinarily or agriculturally (I) salt wherein X is selected from a covalent bond, -C (R5) 2-, -Z- or -C (R5) 2-ZC (R5) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 , CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol , C 1-6 alkylthiol, halogen, CN, C (RS) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 6 -alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1 -6 alkylene) N (C 1-6 alkyl) 2 or a 5 or 6 membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 or R3 and R4 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 Cl -6alkyl, NH2, NHC1-6alkyl or N (C1-6alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the rate of polyolysis of said invertebrate. 4. A method of treating plague infestation comprising inhibiting metamorphosis of an invertebrate by externally contacting said plague with a compound of the formula: <1> formula <1> <1> formula <1> R1 and R1 ' are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3, CO 2 H, CO 2 Cl -6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2 or R1 and R19 taken together are -C (R5) 2-, -C (R5) 2-C (R5) 2 -, -CR 5 = CR 5 -, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol , C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (Ci -6 alkylene) N (C 1-6 alkyl) 2 or a 5 or 6 membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 or R3 and R4 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 Cl -6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the metamorphosis rate of said invertebrate. Plague Infestation Treatment Method according to any one of Claims 1 to 4, characterized in that R 3 and R 3 'are each independently ethyl or methyl. Plague Infestation Treatment Method according to Claim 5, characterized in that R 3 is ethyl. Plague Infestation Treatment Method according to Claim 5, characterized in that R3 is methyl. 8. A plague infestation treatment method according to claim 5 wherein R 3 is methyl. 9. A method of treating plague infestation according to claim 5 wherein R 3 is ethyl. 10. A method of treating plague infestation according to claim 5 wherein R 3 'is ethyl and R 3 is methyl. 11. A plague infestation treatment method according to claim 5 wherein either R 3 is methyl and R 3 is ethyl. 12. A method of treating plague infestation according to any one of claims 1 to 4, wherein both R3 'and R3 are ethyl. A plague infestation treatment method according to any one of claims 1 to 4, characterized in that said compound is a metal chelating agent, wherein the metal chelating agent has at least two polar atoms capable of of simultaneously coordinating with a metal ion, has a clogP value of / 1 and <4; and / or a molar refractivity in the range of 40 to 90 cm 3 / mole. A plague infestation treatment method according to claim 13, wherein the metal chelating agent is not 1,10-phenanthroline. 15. A plague infestation treatment method according to claim 13, wherein the metal chelating agent is not 2,2'-bipyridine. 16. The method of treating plague infestation according to any one of claims 1 to 4, characterized in that said method further comprises contacting said plague with a second pesticide. 17. A method of treating plague infestation comprising reducing the exsheathment of an invertebrate by externally contacting a plague with a compound of formula (II): <formula> formula see original document page 154 < wherein Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN C (R 29) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 1-6 alkyloxy, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R27 is C1-6 alkyl, a branched chain of C1-6 alkyl, C1-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2 or a carbocyclic ring or 5- or 6-membered heterocyclic ring; R28 is hydrogen, C1-6 alkyl or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 1-6 alkenyl, C 1-6 alkynyl chain, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or decrease the exsheathment rate of said invertebrate. 18. A method of treating plague infestation, which comprises decreasing the existence of an invertebrate by externally contacting a plague with a compound of formula (II): wherein: Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN C (R 29) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO 2 H, CO 2 -C 1-6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 27 is C 1-6 alkyl, a C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, C (R 29) 2, N (R 3 O) 2 ring or a carbocyclic ring or ring 5- or 6-membered heterocyclic; R28 is hydrogen, C1-6 alkyl, a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-8 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl chain, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the rate of excision of said invertebrate. 19. A plague infestation treatment method, characterized in that it comprises decreasing the apolysis of an invertebrate by externally contacting a plague with a compound of formula (II): <formula> formula see original document page 157 </formula> in Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 25 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 27 is C 1-6 alkyl, a C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, C (R 29) 2, N (R 30) 2 or a carbocyclic ring or ring 5- or 6-membered heterocyclic; R28 is C1-6 alkyl hydrogen or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl chain, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or slow down the rate of the said invertebrate. 20. A plague infestation treatment method comprising inhibiting metamorphosis of an invertebrate by externally contacting said plague with a compound of the formula: wherein Y1 and Y2 are independently selected from O, NR28 or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R29) 3, CO2 H, CO2 C1-6 alkyl, SO3 H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl, N (C1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 27 is C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, C (R 29) 2, N (R 30) 2 ring or a carbocyclic ring or ring 5- or 6-membered heterocyclic; R28 is hydrogen, C1-6 alkyl, a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl chain, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit or otherwise decrease the metamorphosis rate of said invertebrate. 21. A plague infestation treatment method according to any one of claims 17 to 20, wherein said compound is a metal chelating agent, wherein the metal chelating agent has at least two polar atoms capable of of coordinating simultaneously with a metal ion, has a clogP / 1 value and <4; and / or a molar refractivity in the range of 40 to 90 cm 3 / mole. 22. The method of treating plague infestation according to any one of claims 17 to 20, wherein said method further comprises contacting said plague with a second pesticide. 23. The method of treating plague infestation according to any one of claims 1 to 4 or claims 17 to 20, wherein said method produces a greater decrease in the sheath rate; excystation, apolysis or metamorphosis of what is seen with the administration of 1,10-phenanthroline. 24. An invertebrate plague extermination method, wherein said method comprises contacting said plague with a compound of formula (I): wherein X is selected from a covalent bond, -C (R 5) 2-, -Z- or -C (R 5) 2 -ZC (R 5) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 , CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, halogen, CN, C (Re) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 6 -alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH ( C 1-6 alkylene) N (C 1-6 alkyl) 2 or a 5- or 6-membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 'or R3' and R4 'taken together with the carbon atoms to which they are attached form a 5- or 6-membered carbocyclic or heterocyclic ring; each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 Cl -6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2; each R is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to kill said invertebrate. An invertebrate plague extermination method, wherein said method comprises contacting said plague with a compound of formula (II): wherein Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R29) 3, CO2 H, CO2 C1-6 alkyl, SO3 H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl, N (C1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 27 is C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2 -alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, C (R 29) 2, N (R 30) 2 or a carbocyclic ring or 5- or 6-membered heterocyclic ring; R28 is hydrogen, C1-6 alkyl or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl chain, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to kill said invertebrate. An invertebrate plague extermination method according to claim 24 or 25, wherein said invertebrate is selected from the group consisting of nematodes, trematodes, cestodes, lice, fleas, mites and scabies. , moths, beetles, caterpillar butterflies, termites, arachnids, cockroaches, centipedes, fleas and mites. 27. An invertebrate population remodeling event inhibition method comprising contacting said plague with a compound of formula (I): wherein X is selected from a covalent bond, - C (R5) 2 -, -Z- or -C (R5) 2 -ZC (R5) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 , CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol , C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 6 -alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1 -6 alkylene) N (C 1-6 alkyl) 2 or a 5 or 6 membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 or R3 and R4 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 Cl -6alkyl, NH2, NHC1-6alkyl or N (C1-6alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said invertebrate remodeling event, wherein said invertebrate remodeling event is not egg incubation and said invertebrate is not an ectoparasitic insect. Invertebrate Population Remodeling Event Inhibition Method according to Claim 27, characterized in that R 3 and R 3 'are each independently ethyl or methyl. 29. An invertebrate population remodeling event inhibition method, comprising comprising contacting said plague with a compound of formula (II): wherein Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 1-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29 ) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R27 is C1-6 alkyl, a branched chain of C1-6 alkyl, C1-6 alkenyl, C2-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2 or a carbocyclic ring or 5- or 6-membered heterocyclic ring; R28 is hydrogen, C1-6 alkyl or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl chain, C 1-6 alkenyl, C 1-6 alkynyl, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said invertebrate remodeling event, wherein said invertebrate remodeling event is not egg incubation and said invertebrate is not an ectoparasitic insect. An invertebrate population remodeling event inhibition method according to claim 27 or 29, characterized in that said invertebrate plague is selected from the group consisting of nematodes, trematodes and cestodes. 31. Method of Inhibition of Incubation of Eggs in Non-Ectoparasitic Invertebrates in Invertebrate Population, characterized in that it comprises contacting said invertebrate with a compound of formula (I): <formula> formula see original document page 167 </formula> < wherein X is selected from a covalent bond, -C (RS) 2-, -Z- or -C (RS) 2 -ZC (RS) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 , CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 5) 2-, -C (RS) 2 -C (R 5) 2-, -CR 5 = CR 5 -, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol , C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 6 -alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1 -6 alkylene) N (C 1-6 alkyl) 2 or a 5 or 6 membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 or R3 and R4 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said egg incubation. A method of inhibiting egg incubation in non-ectoparasitic invertebrates in invertebrate populations according to claim 31, wherein R3 and R3 'are each independently ethyl or methyl. 33. Non-Ectoparasitic Invertebrate Egg Incubation Inhibition Method in Invertebrate Population, characterized in that it comprises contacting said invertebrate with a compound of formula (II): <formula> formula see original document page 169 </formula> on whereas Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R29) 3, CO2 H, CO2 C1-6 alkyl, SO3 H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl, N (C1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 27 is C 1-6 alkyl, a C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, C (R 29) 2, N (R 30) 2 or a carbocyclic ring or heterocyclic ring. 5 or 6 members; R28 is hydrogen, C1-6 alkyl or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl chain, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to inhibit said egg incubation. A method of inhibiting egg incubation in non-ectoparasitic invertebrates in an invertebrate population according to claim 31 or claim 33, wherein said invertebrate is selected from the group consisting of nematodes, trematodes and cestodes. A method of inhibiting egg incubation in non-ectoparasitic invertebrates in invertebrate populations according to claim 34, wherein said invertebrate is a nematode. A method of inhibiting egg incubation in non-ectoparasitic invertebrates in an invertebrate population according to claim 35, wherein said nematode is inhibited in its larval phase. 37. An invertebrate plague population control or extermination method, characterized in that it comprises inwardly or externally contacting said plague with a compound of formula (I): wherein X is selected from a covalent bond, - C ( RS) 2-, -Z- or -C (RS) 2-ZC (RS) 2-; R 1 and R 1 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, C (R 6) 3 CO 2 -C 6 -C 6 alkyl, SO 3 H, SO 3 C 6 -C 6 alkyl, NH 2, NHC 1 -C 6 alkyl or N (C 1-6 alkyl) 2 or R 1 and R 19 taken together are -C (R 5) 2-, -C (R 5) 2 -C ( R5) 2-, -CR5 = CR5-, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol , C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 6 -alkyl, NH 2, NHC 1-6 alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1 -6 alkylene) N (C 1-6 alkyl) 2 or a 5 or 6 membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 or R3 and R4 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; each R is independently selected from hydrogen, C1-6 alkyl, a branched C1-6 alkyl, C2-6 alkenyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthiol, CO2H, CO2 C1-4 alkyl, SO3H, SO3C1 chain. -6alkyl, NH2, NHC1-6alkyl or N (C1-6alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to kill or reduce the size of said invertebrate pest population. A method of inhibiting egg incubation in non-ectoparasitic invertebrates in invertebrate populations according to claim 37, wherein R3 and R3 'are each independently ethyl or methyl. 39. An invertebrate plague population control or extermination method, comprising comprising contacting said plague internally or externally with a compound of formula (II): wherein Y1 and Y2 are independently selected from O, NR2S or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 1-6 alkyl, NH 2, NHC 1-6 alkyl, N (C 1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 27 is C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, C (R) 2, N (R 30) 2 or a carbocyclic ring or 5- or 6-membered heterocyclic ring; R28 is hydrogen, C1-6 alkyl or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched chain of C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof in an amount effective to kill or reduce the size of said invertebrate pest population. 40. Chelating Agent Selection Method As Candidate Invertebrate Remodeling Event Inhibitor From Collection of Metal Chelating Agents, characterized in that said method comprises selecting a metal chelating agent having at least two polar atoms. able to coordinate simultaneously with a metal ion and i. a clogP / 1 value and <4; and / or ii. molar refractivity in the range of 40 to 90 cm3 / mole. 41. Chelating Agent Selection Method As Candidate Inhibitor of Invertebrate Remodeling Event from Collection of Metal Chelating Agents, characterized in that said chelating agent is of formula (II): where Y1 and Y2 are independently selected from O, NR28 or S; R 21, R 22, R 23 and R 24 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R29) 3, CO2 H, CO2 C1-5 alkyl, SO3H, SO5 C1-6 alkyl, NH2, NHC1-6 alkyl, N (C1-6 alkyl) 2 or a carbocyclic or heterocyclic ring; or R21 and R22 or R22 and R23 and R24 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R 25 and R 26 are independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthio, halogen, CN, C (R 29) 3, CO2H, CO2 C1-6 alkyl, SO3H, SO3 C1-6 alkylNH2, NHC1-6 alkyl or N (C1-6 alkyl) 2; or R 25 and R 26 together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; R27 is C1-6 alkyl, a branched C1-6 alkyl, C2-6alkenyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, C (R29) 2, N (R30) 2 or a carbocyclic ring or heterocyclic ring of 5 or 6 members; R28 is hydrogen, C1-6 alkyl or a branched C1-6 alkyl chain; R29 is hydrogen or halogen; and each R 30 is independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl chain, C 2-6 alkenyl, C 2-6 alkynyl, a 5- or 6-membered carbocyclic ring or heterocyclic ring; or a pharmaceutically, veterinarily or agriculturally acceptable salt thereof; said method comprising selecting a metal chewing agent of formula (II) having at least two polar atoms capable of coordinating simultaneously with a metal ion and i) a clogP / 1 and <4 value; and / or ii) a molar refractivity in the range of 40 to 90 cm 3 / mole. 42. Chelating Agent Selection Method As Candidate Inhibitor of Invertebrate Remodeling Event from Collection of Metal Chelating Agents, characterized in that said chelating agent is of formula (I): <formula> formula see original where X is selected from a covalent bond, -C (R5V, -Z- or -C (R5) 2 -ZC (RS) 2-; R1 and R1 'are independently selected from from hydrogen, C1-6 alkyl, a branched chain C1-6 alkyl, C2-6 alkenyl, C1-6 alkynyl, hydroxy, C1-6 alkoxy, thiol, C1-6 alkylthio, halogen, C (R6) 3, CO2H, CO2 C1-6 alkyl, SO3H , C03 -C6 alkyl, NH2, NHC1-6 alkyl or N (C1-6 alkyl) 2 or R1 and R19 taken together are -C (R5) 2-, -C (R5) 2 -C (R5) 2-, -CR5 = CR 5 -, C (O), C (S) or NH; R 2, R 2 ', R 3, R 3', R 4 and R 4 'are independently selected from hydrogen, C 1-6 alkyl, a branched C 1-6 alkyl chain C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, halogen, CN, C (R 6) 3, CO 2 H, CO 2 -C 6 alkyl, SO 3 H, SO 3 C 6 -alkyl, NH 2, NHC 1 -alkyl or N (C 1-6 alkyl) 2, -CH 2 CHNH (CO 2 H), NH (C 1-5 alkylene) N (C 1-6 alkyl) 2 or a 5 or 6 membered carbocyclic or heterocyclic ring; or R2 and R3 or R3 and R4 and / or R2 and R3 or R3 and R4 taken together with the carbon atoms to which they are attached form a 5 or 6 membered carbocyclic or heterocyclic ring; each R 5 is independently selected from hydrogen, C 1-6 alkyl, a branched chain C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, hydroxy, C 1-6 alkoxy, thiol, C 1-6 alkylthiol, CO 2 H, C02 -C 1-6 alkyl, SO 3 H, SO 3 Cl -6 alkyl, NH2, NHC1-5 alkyl or N (C1-6 alkyl) 2; each R 6 is independently selected from hydrogen and halogen; and Z is selected from a covalent bond, -NH-, -O-, -S-, -C (O) - and -C (S) -; said method comprising selecting a metal chewing agent of formula (I) having at least two polar atoms capable of coordinating simultaneously with a metal ion and i) a clogP / 1 value and <4; and / or ii) a molar refractivity in the range of 40 to 90 cc / mole.